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Статті в журналах з теми "MSC culture":
1
Peters, R. E., M. Heikenwälder, and A. Knuth. "Expansion of umbilical cord blood mesenchymal stem cells." Journal of Clinical Oncology 27, no. 15_suppl (May 20, 2009): 7103. http://dx.doi.org/10.1200/jco.2009.27.15_suppl.7103.
Анотація:
7103 Background: Umbilical cord blood (UCB) is known to harbor 2 major types of stem cells, the hematopoietic stem cells (HSC) & the non-hematopoietic or mesenchymal stem cells (MSC). Under appropriate conditions, MSCs can give rise to cells of bone, fat, hepatic lineages, etc. Based on this potential, MSC hold promise for clinical applications in regenerative medicine. Methods: Stroma-free liquid culture: UCB cryopreserved mononuclear cells (MNC) were cultured in the presence of early growth factors: Flt-3 & SCF (25ng/ml), MGDF (10ng/ml) & human serum (10%). MNC derived adherent MSC were passaged at day 14 during HSC expansion & after enriching in MesenCult medium. Results: We developed a technology to generate & expand HSC & stromal/ MSC from all UCB units (5/5) at the same time using one culture system (stroma-free liquid culture). Following repeated passages, MSC count increased 357- 600-folds & CFU-Fibroblasts colonies (CFU-F) increased too (61–513 & 648–697) after 10 and 20 passages respectively. We used the CFU-F assay to demonstrate MSC activity in stromal cell formation in vitro. Phenotypically, MSC were negative for hematopoietic antigens (CD45, CD34 & CD14) & MHC class-II but >95% + for CD73, CD105, CD29, CD44 & MHC class I. To demonstrate MSC differentiation capacity in vitro, cells were incubated in various induction media to differentiate into adipocytes (fat)), osteoblasts (bone) and hepatocytes (liver) at passage 5. Following induction, positive staining with oil red O for cells of adipocyte and with alkaline phosphatase for cells of osteoblsts lineages was observed. The identity of hepatocytes was verified by the characteristic hexagonal hepatocytic shape as well as albumin, cytokeratin (CK) 18 and CK14 expression, as assessed by flow cytometry. Our data were corroborated by RT-PCR analysis. Conclusions: MSC described herein exhibit in vitro properties of multipotent stem cells. The established, stroma-free culture system facilitates expansion of MSC from all tested UCB units. Our data underline that it will be possible in the future to substitutes properly differentiated hepatocytes which might lead to efficient applications in patients suffering from various end stage liver disease. No significant financial relationships to disclose.
2
Prewitz, Marina, Friedrich Philipp Seib, Martin Bornhaeuser, and Carsten Werner. "Engineering Biomimetic Culture Systems: Impact On Human Bone Marrow-Derived Stem Cells." Blood 114, no. 22 (November 20, 2009): 3628. http://dx.doi.org/10.1182/blood.v114.22.3628.3628.
Анотація:
Abstract Abstract 3628 Poster Board III-564 The bone marrow (BM) harbours haematopoietic stem/progenitor cells (HSCs) in anatomically distinct sites (niches) where HSCs are subjected to regulatory cues such as cytokines, cell-cell contacts and extra-cellular matrix (ECM) all of which control stem cell fate. In particular mesenchymal stromal cells (MSCs) are an integral part of the bone marrow and are known to be key regulators of the HSC niche. We have previously shown that bio-artificial scaffolds can have a significant impact on the in vitro behaviour of MSCs. Here, we are therefore focussing on the role of (native) ECM within the MSC-HSC microenvironment by building on our previous findings and published data (Seib et al.,Tissue Eng Part A., 2009 in press). Thus the aim of the current study is (a) to identify niche-specific ECM components and (b) the use of such ECMs for in vitro culture of BM-derived stem cells. To mimic the natural ECM composition of the BM, different ECM types were generated from BM-derived cells using (a) Dexter cultures, (b) standard MSC cultures, (c) MSCs subjected to osteogenic differentiation. After 10 days of culture those MSC-derived ECMs were decellularised using 0.5% Triton-X and 20mM NH4OH leaving only the ECM behind (verified by scanning electron microscopy). Those ECMs were used as a substrate for a second culture of MSCs, which were analysed for their proliferation and differentiation potential. Cell-free ECM from standard MSC cultures improved MSC proliferation compared to cells grown on regular tissue culture plastic (TCP) over the period of 8 days. Most notably, all cell-free ECM preparations lead to a significant difference in the cytoskeletal arrangement of MSCs during the first 2 days of culture compared to TCP controls. Cultivation of MSCs on native ECM provided a guiding structure for those cells to grow into, and helped to maintain an elongated cell shape compared to substantial cell spreading on TCP (roundness 0.2 versus 0.5 and cell area of 2.2 versus 8.2mm2, respectively, p<0.001, n=60. A factor of 1 was set to equate to a perfect circle). Next, we investigate if native ECM could either directly improve HSC cultures or maximise MSC feeder characteristics. For the latter set of studies MSCs were initially cultured for 7 days on cell-free ECM (from standard MSC cultures) and subsequently co-cultured with human peripheral blood CD34+ HSCs in serum free medium supplemented with cytokines (Tpo, Flt3, and SCF at 10ng/ml). Following a 14 day culture period up to 3.5-fold more CD34+ cells were present in ECM co-cultures compared to TCP co-cultures that was accompanied with an overall expansion of CD45+ cells of 109-fold versus 35-fold, respectively. Our data suggest that ECM preparations derived from MSCs might be useful to accomplish better expansion of HSCs under defined culture conditions. In addition, this system permits the identification of bimolecular key components that can be utilized in the future design of simple and robust carrier systems for improved HSC maintenance in vitro. Figure HSC-MSC co-culture on preformed ECM substrates. (A) MSC-derived ECM (from standard MSC culture) following cell lysis (complete absence of cells). (B) Growth of a new set of MSCs on ECM substrates as shown in (A). (C) HSC-MSC co-culture on ECM substrates. Scale bars at 2μm. Arrow heads point out ECM structures. Figure HSC-MSC co-culture on preformed ECM substrates. (A) MSC-derived ECM (from standard MSC culture) following cell lysis (complete absence of cells). (B) Growth of a new set of MSCs on ECM substrates as shown in (A). (C) HSC-MSC co-culture on ECM substrates. Scale bars at 2μm. Arrow heads point out ECM structures. Disclosures: No relevant conflicts of interest to declare.
3
Robinson, Simon N., Paul J. Simmons, Nathalie Brouard, Shannon Kidd, Hong Yang, William K. Decker, Dongxia Xing, et al. "Efficacy of ‘Off-the-Shelf’, Commercially-Available, Third-Party Mesenchymal Stem Cells (MSC) in Ex Vivo Cord Blood (CB) Co-Culture Expansion." Blood 110, no. 11 (November 16, 2007): 4106. http://dx.doi.org/10.1182/blood.v110.11.4106.4106.
Анотація:
Abstract INTRODUCTION: Our previous studies have shown that clinically-relevant levels of hematopoietic stem and progenitor cell (HSPC) expansion are possible by ex vivo co-culture of cord blood (CB) mononuclear cells (MNC) with third-party bone marrow (BM)-derived mesenchymal stem cells (MSC) and growth factors.1 A recently activated M. D. Anderson protocol requires that BM from a haplo-identical family member be used for the de novo generation of sufficient MSC for subsequent co-culture, a process requiring ∼3 weeks. Time constraints, uncertainties associated with the identification of a suitable BM donor and potential variation in MSC performance make logistical execution of this strategy difficult. We therefore investigated the potential efficacy of ‘off-the-shelf’ commercially-available sources of MSC. Since MSC do not express HLA-II (DR) they are non-immunogenic, suggesting that this might be a valuable alternative strategy. We compared ex vivo CB HSPC expansion obtained following CB MNC co-culture with 2 commercially-available research-grade MSC isolated by density separation and plastic adherence (MSC#1, Cambrex, Walkersville, MD and MSC#2, Allcells, Emeryville, CA). A third MSC, isolated by Stro-12 selection (MSC#3, supplied by PJS) was also evaluated. METHODS: Two MDACC frozen CB units (CB#1&2) were thawed, washed and co-cultured with adherent monolayers from each MSC. Total nucleated cell (TNC) and HSPC (CD34+ cells and colony-forming units, CFU) numbers were measured at input (Day 0) and output (Day 14). RESULTS: TNC and HSPC numbers revealed that the 2 commercially-available research-grade MSC (MSC#1&2) supported ex vivo CB HSPC expansion. MSC TNC CB34+ CFU n/a - not available CB#1 #1 x 6 x23 n/a #2 x 3 x 8 x15 #3 x 6 x16 x23 CB#2 #1 x 7 x20 x31 #2 x 5 x10 x20 #3 x10 x16 x34 1 Robinson et al. x13 x14 x25 MSC#2 performed less well than MSC#1 for both CB units suggesting that variation may exist between individual MSC. These data suggest that the screening of clinical-grade MSC that perform optimally during ex vivo expansion co-culture might be warranted to best utilize this ‘off-the-shelf’ strategy. Data were similar to previous reports where TNC, CD34+ and CFU numbers were shown to increase approximately 13, 14 and 25-fold, respectively.1 Data were also similar for MSC#3, suggesting that the method used to isolate MSC does not appear to be an important variable for effective CB MNC/MSC co-culture. CONCLUSION: Although research-grade MSC were compared from different commercial sources, these data suggest that, in principle, commercially-available clinical-grade MSC might prove a valuable ‘off-the-shelf’ option, potentially reducing the time to therapy and addressing concerns associated with identifying a BM donor and variation in MSC performance. Future studies will evaluate FDA-compliant MSC that could be used clinically.
4
Bucher, Christian, Amiq Gazdhar, Lorin M. Benneker, Thomas Geiser, and Benjamin Gantenbein-Ritter. "Nonviral Gene Delivery of Growth and Differentiation Factor 5 to Human Mesenchymal Stem Cells Injected into a 3D Bovine Intervertebral Disc Organ Culture System." Stem Cells International 2013 (2013): 1–9. http://dx.doi.org/10.1155/2013/326828.
Анотація:
Intervertebral disc (IVD) cell therapy with unconditioned 2D expanded mesenchymal stem cells (MSC) is a promising concept yet challenging to realize. Differentiation of MSCs by nonviral gene delivery of growth and differentiation factor 5 (GDF5) by electroporation mediated gene transfer could be an excellent source for cell transplantation. Human MSCs were harvested from bone marrow aspirate and GDF5 gene transfer was achieved byin vitroelectroporation. Transfected cells were cultured as monolayers and as 3D cultures in 1.2% alginate bead culture. MSC expressed GDF5 efficiently for up to 21 days. The combination of GDF5 gene transfer and 3D culture in alginate showed an upregulation of aggrecan and SOX9, two markers for chondrogenesis, and KRT19 as a marker for discogenesis compared to untransfected cells. The cells encapsulated in alginate produced more proteoglycans expressed in GAG/DNA ratio. Furthermore, GDF5 transfected MCS injected into an IVD papain degeneration organ culture model showed a partial recovery of the GAG/DNA ratio after 7 days. In this study we demonstrate the potential of GDF5 transfected MSC as a promising approach for clinical translation for disc regeneration.
5
Tormin, Ariane, Jan Claas Brune, Stuart Walsh, Johan Richter, Xiaolong Fan, and Stefan Scheding. "Human Primary Mensenchymal Stromal Progenitor Cells Are Highly Enriched in Both, the CD271+/CD146+ and CD271+/CD146− Bone Marrow Population with the Latter Acquiring CD146 Expression upon Culture in-Vitro." Blood 112, no. 11 (November 16, 2008): 2422. http://dx.doi.org/10.1182/blood.v112.11.2422.2422.
Анотація:
Abstract Culture-derived mesenchymal stromal cells (MSC), which are attractive candidates for clinical cell therapy approaches, arise from primary MSC progenitor/stem cells in the bone marrow. Recently, several groups have reported markers (CD271, CD146, GD2, SSEA4, etc.) that allowed for an enrichment of CFU-F, i.e. primary MSC progenitors. However, the exact phenotype of the bona fide mesenchymal stem/progenitor cells has not yet not been sufficiently defined. We therefore aimed to investigate primary MSC in bone marrow subpopulations defined by the expression of CD271 and CD146, as both markers have been reported to contain all assayable CFU-F and stromal stem cells, respectively (Quirici et al., Exp. Hematol. 2002; Sacchetti et al, Cell, 2007). Utilizing multi-color flow cytometry, unfractionated human bone marrow mononuclear cells (BM-MNC) were found to contain 0.05 ± 0.05% CD271+/CD146+ cells, whereas 0.82 ± 0.43% and 0.71 ± 0.23% were single-positive for CD271 and CD146, respectively. CD271/CD146 subpopulations were FACS sorted from lineage-depleted BM-MNC (RosetteSep) and assayed for CFU-F content (n=3). CFU-F could not be detected in the CD271−/CD146− fraction. In contrast, CFU-F initiating cells were highly enriched in the CD271+/CD146+/CD45−/low fraction (1.1 ± 0.2 CFU-F per 10 plated cells), which corresponds to a ca. 400-fold enrichment compared to the entire lineage-depleted fraction (2.7 ± 3.4 CFU-F per 1 × 104 plated cells). Of note, CFU-F could also be assayed at high frequency from CD271+/CD146− cells (20.4 ± 22.6 CFU-F per 1 × 104 plated cells). Generally, CFU-F were not found in the CD271+/CD146+/CD45+ and the CD271−/CD146+ fractions, and were also not detectable within the whole CD271+/CD45+ population of unfractionated BM-MNC (n=4), which, however, gave rise to erythropoietic colonies. The two CFU-F enriched populations, i.e. CD271+/CD146+/CD45−/low and CD271+/CD146− cells, were then cultured under standard MSC growth conditions. MSC derived from both populations exhibited a typical MSC surface marker profile (CD105+, CD90+, CD73+, HLA-class I+, CD45−, CD34−, CD19−, CD14−, HLA-DR−) and typical MSC differentiation (adipocytes, osteoblasts, chondrocytes). Interestingly, MSC generated from CD271+/CD146− cells became positive for CD146 in culture and stable CD146 expression over time was observed for MSC from both populations (up to the 5th passage, average 82 ± 11%). In contrast, over the same culture period CD271 expression decreased with passage number and an average of only 10 ± 4% of the cultured cells remained positive for CD271. To further characterize the CFU-F enriched subpopulations, single cells from CD271+/CD146+/CD45−/low and CD271+/CD146−/CD45−/low cells were sorted into fibronectin-coated 96-well plates to investigate colony growth and differentiation potential. CFU-F frequencies in this assay were 4 per 96 seeded cells for both populations and all but one of the CD271+/CD146+/CD45−/low clones could be further expanded in culture. Subpopulation-derived clones were capable of typical MSC differentiation and MSC derived from CD271+/CD146−/CD45−/low clones–similar to the bulk cultures–became CD146 positive (89 ± 12%) after 2 passages, whereas here CD271 expression was not lost. Taken together, CD271+/CD146+/CD45−/low and CD271+/CD146−/CD45−/low bone marrow cells are highly enriched for primary MSC progenitor cells. The difference in CD146 expression, which disappears in culture, might relate different localizations of the primary cells in the marrow but might possibly also reflect functional differences, e.g. in stemness. Accordingly, experiments addressing in-situ location, in vivo differentiation potential, gene expression and surface-marker expression profiling of primary MSC are currently under way.
6
Giallongo, Cesarina, Daniele Tibullo, Nunziatina Laura Parrinello, Piera La Cava, Giuseppina Camiolo, Nunzia Caporarello, Carmelina Daniela Anfuso, et al. "Mesenchymal Stem Cells (MSC) from Patients with Multiple Myeloma Promote Myeloid Cells to Become Granulocytic-Myeloid-Derived Suppressor Cells (G-MDSC) with Immunosuppressive, Bone Resorption and Pro-Angiogenic Activity." Blood 128, no. 22 (December 2, 2016): 4458. http://dx.doi.org/10.1182/blood.v128.22.4458.4458.
Анотація:
Abstract Purpose A well-recognized feature of MM is the intimate relationship between plasma cells and bone marrow microenvironment, which is mainly composed of MSC, endothelial cells, immune cells and extracellular matrix. G-MDSC accumulate in the tumor microenvironment during tumor development promoting tumor growth and immunosuppression. Aim Analyzing MSC from MGUS, Smoldering myeloma (SMM) and MM patients in promoting G-MDSC generation. Methods Human peripheral blood mononucleated cells (PBMC) isolated from healthy subjects (HS) were cultured alone and with HS- (n=10), MGUS- (n=10), SMM- (n=4) or MM-MSC (n=14)(1:100). After 6 days, G-MDSC were isolated using anti-CD66b magnetic microbeads and the phenotype(CD11b+CD33+CD14-HLADR-) was confirmed by cytofluorimetric analysis. Results Only G-MDSC educated by SMM- and MM-MSC co-cultures (MSCed-G-MDSC) were able to suppress T cell proliferation when cultured with normal lymphocytes (p<0.001) compared to G-MDSC control (isolated from PBMC cultured in medium alone). SMM- and MM-MSCed-G-MDSC significantly up-regulated Arg1, NOS2, TNFα and CEBPA, a transcription factor promoting suppressive phenotype. Since also the angiogenic factor BV8 was significantly up-regulated, we next investigated the pro-angiogenic effect in vitro co-culturing MSCed-G-MDSC with Human Brain Microvascular Endothelial Cells (HBMEC) (1:2). After 5 h, we observed that MM-MSCed-G-MDSC were able to increase both tube length and number of branch points compared to G-MDSC control (p<0.05). Moreover, MM-MSCed-G-MDSC were able to digest bone matrix in vitro (p<0.01). Adding Bortezomib (5 nM), Lenalidomide (10 μM) or Pomalidomide (1 nM) during co-culture with MM-MSC, isolated G-MDSC showed a significant reduction of pro-angiogenic and bone resorption activity (p<0.05) but did not lose immunosuppressive ability. Conclusion MSC play a key role promoting tumor microenvironment transformation in SMM and MM patients.Indeed, only SMM- and MM-MSC and not MGUS-MSC are able to activate myeloid cells in G-MDSC with immunosuppressive, pro-angiogenic, and bone resorptionactivity. Disclosures No relevant conflicts of interest to declare.
7
Maijenburg, Marijke W., Marion Kleijer, Kim Vermeul, Erik P. J. Mul, Floris P. J. van Alphen, C. Ellen van der Schoot, and Carlijn Voermans. "Primary Bone Marrow-Derived MSC Subsets Have Distinct Wnt-Signatures Compared to Conventionally Cultured MSC and Differ in Their Capacity to Support Hematopoiesis in Vitro,." Blood 118, no. 21 (November 18, 2011): 3414. http://dx.doi.org/10.1182/blood.v118.21.3414.3414.
Анотація:
Abstract Abstract 3414 Mesenchymal stromal cells (MSC) are of promising therapeutic use to suppress immunogenic responses following transplantation, and to support expansion of hematopoietic stem- and progenitors cells (HSPC) from small transplants derived for instance from cord blood. Culture-expanded MSC produce a wide variety and quantity of Wnt-proteins and the crucial role of Wnt-signaling in the hematopoietic stem cell niche is well established. However, studies addressing Wnt-signaling in MSC have (i) only focused on culture-expanded MSC and (ii) did not discriminate between phenotypically distinct subpopulations which are present in bulk cultures of expanded MSC. Recently we identified three new subpopulations of MSC in human bone marrow (BM) based on expression of CD271 and CD146: CD271brightCD146−, CD271brightCD146+, CD271−CD146+. These fractions co-express the “classical” MSC markers CD90 and CD105 and lack expression of CD45 and CD34 (Maijenburg et al, Blood 2010, 116, 2590). We and others demonstrated that the adult BM-derived CD271brightCD146− and CD271brightCD146+ cells contain all colony forming units-fibroblasts (Maijenburg et al, Blood 2010, 116, 2590; Tormin et al, Blood 2010, 116, 2594). To investigate how these primary subsets functionally compare to conventional, culture-expanded MSC, we investigated their Wnt-signature and hematopoietic support capacity. To this end, we sorted CD271brightCD146− and CD271brightCD146+ cells from human adult BM (n=3) and compared their Wnt-signatures obtained by Wnt-PCR array to the profiles from cultured MSC from the same donors. Fifteen genes were consistently differentially expressed in the two sorted uncultured subsets compared to their conventionally cultured counterparts. Expression of CCND1, WISP1 and WNT5B was strongly increased, and WNT5A was only detected in the conventionally cultured MSC. In contrast, WNT3A was exclusively expressed by sorted primary CD271brightCD146− and CD271brightCD146+ cells, that also expressed higher levels of JUN, LEF1 and WIF1. The differences in Wnt (target)-gene expression between CD271brightCD146− and CD271brightCD146+ cells were more subtle. The Wnt-receptors LRP6 and FZD7 were significantly higher expressed in CD271brightCD146+ cells, and a trend towards increased expression in the same subset was observed for CTNNB1, WNT11 and MYC. When the sorted subsets were cultured for 14 days (one passage), the differences in Wnt-related gene expression between the subsets was lost and the expanded sorted cells acquired an almost similar Wnt-signature as the MSC cultured from BM mononuclear cells from the same donors. The cultured subsets lost the expression of Wnt3a and gained the expression of Wnt5a, similar to the unsorted MSC cultured from the same donors in parallel. Despite the loss of a distinct Wnt-signature, co-culture experiments combining the sorted MSC subsets with human HSPC revealed that CD271brightCD146+ cells have a significantly increased capacity to support HSPC in short-term co-cultures (2 weeks) compared to CD271brightCD146− cells (p<0.021, n=3), which was analyzed in hematopoietic colony assays following co-culture. In contrast, a trend towards better long-term hematopoietic support (co-culture for 6 weeks) was observed on CD271brightCD146− cells. In conclusion, we demonstrate for the first time that primary sorted uncultured MSC subsets have a distinct Wnt-signature compared to cultured unsorted MSC and display differences in hematopoietic support. As it was recently shown that CD271brightCD146− and CD271brightCD146+ MSC localize to separate niches in vivo (Tormin et al, Blood 2011), our data indicate that the two MSC subsets are not necessarily distinct cell types and that the different Wnt-signature may be a reflection of these distinct microenvironments. Cell culturing for only one passage dramatically changed the Wnt-signature of the sorted MSC subsets, indicating that Wnt-signaling in in vitro expanded MSC does not resemble the Wnt-signature in their tissue resident counterparts in vivo. Disclosures: No relevant conflicts of interest to declare.
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Burk, Janina, Anna Sassmann, Cornelia Kasper, Ariane Nimptsch, and Susanna Schubert. "Extracellular Matrix Synthesis and Remodeling by Mesenchymal Stromal Cells Is Context-Sensitive." International Journal of Molecular Sciences 23, no. 3 (February 3, 2022): 1758. http://dx.doi.org/10.3390/ijms23031758.
Анотація:
Matrix remodeling could be an important mode of action of multipotent mesenchymal stromal cells (MSC) in extracellular matrix (ECM) disease, but knowledge is limited in this respect. As MSC are well-known to adapt their behavior to their environment, we aimed to investigate if their mode of action would change in response to healthy versus pathologically altered ECM. Human MSC-derived ECM was produced under different culture conditions, including standard culture, culture on Matrigel-coated dishes, and stimulation with the pro-fibrotic transforming growth factor-β1 (TGFβ1). The MSC-ECM was decellularized, characterized by histochemistry, and used as MSC culture substrate reflecting different ECM conditions. MSC were cultured on the different ECM substrates or in control conditions for 2 days. Culture on ECM increased the presence of surface molecules with ECM receptor function in the MSC, demonstrating an interaction between MSC and ECM. In MSC cultured on Matrigel-ECM and TGFβ1-ECM, which displayed a fibrosis-like morphology, gene expression of collagens and decorin, as well as total matrix metalloproteinase (MMP) activity in the supernatant were decreased as compared with control conditions. These results demonstrated that MSC adapt to their ECM environment, which may include pathological adaptations that could compromise therapeutic efficacy.
9
Koh, Benson, Nadiah Sulaiman, Mh Busra Fauzi, Jia Xian Law, Min Hwei Ng, Too Lih Yuan, Abdul Ghani Nur Azurah, Mohd Heikal Mohd Yunus, Ruszymah Bt Hj Idrus, and Muhammad Dain Yazid. "A Three-Dimensional Xeno-Free Culture Condition for Wharton’s Jelly-Mesenchymal Stem Cells: The Pros and Cons." International Journal of Molecular Sciences 24, no. 4 (February 13, 2023): 3745. http://dx.doi.org/10.3390/ijms24043745.
Анотація:
Xeno-free three-dimensional cultures are gaining attention for mesenchymal stem cell (MSCs) expansion in clinical applications. We investigated the potential of xeno-free serum alternatives, human serum and human platelet lysate, to replace the current conventional use of foetal bovine serum for subsequent MSCs microcarrier cultures. In this study, Wharton’s Jelly MSCs were cultured in nine different media combinations to identify the best xeno-free culture media for MSCs culture. Cell proliferation and viability were identified, and the cultured MSCs were characterised in accordance with the minimal criteria for defining multipotent mesenchymal stromal cells by the International Society for Cellular Therapy (ISCT). The selected culture media was then used in the microcarrier culture of MSCs to determine the potential of a three-dimensional culture system in the expansion of MSCs for future clinical applications, and to identify the immunomodulatory potential of cultured MSCs. Low Glucose DMEM (LG) + Human Platelet (HPL) lysate media appeared to be good candidates for replacing conventional MSCs culture media in our monolayer culture system. MSCs cultured in LG-HPL achieved high cell yield, with characteristics that remained as described by ISCT, although the overall mitochondrial activity of the cells was lower than the control and the subsequent effects remained unknown. MSC microcarrier culture, on the other hand, showed comparable cell characteristics with monolayer culture, yet had stagnated cell proliferation, which is potentially due to the inactivation of FAK. Nonetheless, both the MSCs monolayer culture and the microcarrier culture showed high suppressive activity on TNF-α, and only the MSC microcarrier culture has a better suppression of IL-1 secretion. In conclusion, LG-HPL was identified as a good xeno-free media for WJMSCs culture, and although further mechanistic research is needed, the results show that the xeno-free three-dimensional culture maintained MSC characteristics and improved immunomodulatory activities, suggesting the potential of translating the monolayer culture into this culture system in MSC expansion for future clinical application.
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Kaminska, Agnieszka, Aleksandra Wedzinska, Marta Kot, and Anna Sarnowska. "Effect of Long-Term 3D Spheroid Culture on WJ-MSC." Cells 10, no. 4 (March 24, 2021): 719. http://dx.doi.org/10.3390/cells10040719.
Анотація:
The aim of our work was to develop a protocol enabling a derivation of mesenchymal stem/stromal cell (MSC) subpopulation with increased expression of pluripotent and neural genes. For this purpose we used a 3D spheroid culture system optimal for neural stem cells propagation. Although 2D culture conditions are typical and characteristic for MSC, under special treatment these cells can be cultured for a short time in 3D conditions. We examined the effects of prolonged 3D spheroid culture on MSC in hope to select cells with primitive features. Wharton Jelly derived MSC (WJ-MSC) were cultured in 3D neurosphere induction medium for about 20 days in vitro. Then, cells were transported to 2D conditions and confront to the initial population and population constantly cultured in 2D. 3D spheroids culture of WJ-MSC resulted in increased senescence, decreased stemness and proliferation. However long-termed 3D spheroid culture allowed for selection of cells exhibiting increased expression of early neural and SSEA4 markers what might indicate the survival of cell subpopulation with unique features.
Дисертації з теми "MSC culture":
1
Maillot, Charlotte. "Quantification and impact of microcarrier collisions during mesenchymal stem cell culture in bioreactors." Electronic Thesis or Diss., Université de Lorraine, 2022. http://www.theses.fr/2022LORR0314.
Анотація:
Les thérapies cellulaires et tissulaires sont des thérapies innovantes à base de cellules du patient (on parle alors de thérapies autologues) ou de cellules d'un donneur compatible (on parle alors de thérapies allogéniques). Contrairement aux médicaments chimiques ou à base des protéines souvent recombinantes (anticorps monoclonaux par exemple), le fait d'utiliser des cellules en tant que produit permet d'envisager de nouvelles cibles thérapeutiques qui peuvent être difficiles d'accès, et aussi d'augmenter la spécificité en engageant une transition vers des médecines ciblées et personnalisées. Cependant, le caractère vivant des thérapies (pour lesquelles les cellules en elles-mêmes constituent la thérapie) amène aussi une technicité en terme de production. En effet, les processus de production de ces nouvelles médecines doivent garantir une qualité des produits cellulaires dont les attributs et moyens de vérification d'activité sont souvent peu définis et/ou complexes. C'est d'autant plus une problématique que la qualité des matières premières peut être variable d'un donneur à un autre, particulièrement dans le cas de thérapies autologues pour lesquelles les patients ont bien souvent un système immunitaire et une qualité de don affecté par leur maladie. Pour commencer, une approche quantitative pour estimer la croissance cellulaire et la cinétique de mort causées par les collisions microporteur-microporteur dans les flacons Erlenmeyer spinner est décrite. Pour cela, les cellules ont été cultivées à différentes concentrations de microporteurs en utilisant deux types de microporteurs : Cytodex-1 et Synthemax II. Des cultures complémentaires ont été réalisées en ajoutant diverses concentrations de particules de même taille et densité que les microporteurs en vue de fournir des informations spécifiques sur la façon dont des particules supplémentaires peuvent avoir un impact sur la croissance des MSC sur les microporteurs. De plus, des éléments de caractérisation MSC ont été réalisés pour ces expériences afin de comprendre non seulement l'impact des interactions microporteur-microporteur sur la croissance mais aussi sur des éléments définis de la qualité cellulaire. En parallèle, afin d'estimer la quantité et l'intensité des collisions microporteurs-microporteurs dans une géométrie spécifique, des expériences ont été réalisées en utilisant à la fois l'atténuation de la lumière par les microporteurs Cytodex-1 (pour estimer la concentration locale de microporteurs) et des signaux acoustiques qui proviennent de particules entrant en collision avec un hydrophone (pour estimer la fréquence et l'intensité de la collision microporteur-capteur). Ces expériences ont fourni des éléments pour estimer la quantité de collisions de particules que les MSC peuvent percevoir pendant des phases dynamiques et stables, spécifiques aux cultures cellulaires en bioréacteur. Enfin, une approche basée sur les bioréacteurs pour la fabrication de MSC est présentée en se concentrant sur les aspects biologiques de l'impact de la concentration et de l'agitation des particules sur la croissance et les attributs de qualité des MSC. Pour cela, diverses cultures de MSC ont été réalisées dans des STR avec des concentrations de particules et des stratégies d'agitation variables. Les MSC produites dans ces conditions ont ensuite été caractérisées pour définir si certains attributs de qualité pouvaient être affectés par des paramètres tels que la concentration et/ou l'agitation des microporteursTo date, bottlenecks persist concerning deep scientific understanding of how various process parameters will impact the Mesenchymal Stem Cell production. Specifically applied to microcarrier-based expansion processes of WJ-MSC's, very little information is available to characterize the impact of microcarrier concentration on MSC growth and death rates or on critical quality attributes which may have crucial and possibly dangerous clinical impacts. As a result, the following work proposes to rationally describe the impact of particle concentration on MSC growth through a pluri-disciplinary characterization of microcarrier-microcarrier interactions in agitated conditions. In order to do so the biological and physical aspects of this work will be presented. To begin with, a quantitative approach to estimate cell growth and death kinetics caused by microcarrier-microcarrier collisions in both Erlenmeyer Flasks and Spinner Flasks is described. For this, cells were grown at various microcarrier concentrations using two microcarrier types : Cytodex-1 and Synthemax II. Complementary cultures were performed by adding various concentrations of particles with the same size and density as microcarriers in view of providing specific information on how additional particles may impact MSC growth on microcarriers. In addition, elements of MSC characterization were performed for these experiments to understand not only the impact of microcarrier-microcarrier interactions on growth but also on defined elements of cell quality. In parallel, in order to estimate the amount and intensity of microcarrier-microcarrier collisions in a specific tank geometry, experiments were performed using both the attenuation of light by Cytodex-1 microcarriers (to estimate local microcarrier concentration) and the acoustic signal which comes from particles colliding with a hydrophone (to estimate microcarrier-sensor collision frequency and intensity). These experiments provided elements to estimate the amount of particle collisions that MSC's may perceive during specific dynamic and steady phases of cell culture in STR's. Lastly, a bioreactor-based approach to MSC manufacturing will be presented focusing on biological aspects of how particle concentration and agitation impacts MSC growth and quality attributes. For this, various MSC cultures were performed in STRs with varying particle concentrations and agitation strategies. The MSC's produced in these conditions were then characterized to define if certain critical quality attributes could be affected by parameters such as microcarrier concentration and/or agitation
2
Futrega, Katarzyna. "Device and application development for haematopoietic stem and progenitor cell (HSPC) and mesenchymal stromal cell (MSC) 3D spheroid cultures." Thesis, Queensland University of Technology, 2016. https://eprints.qut.edu.au/92605/1/Katarzyna_Futrega_Thesis.pdf.
Анотація:
With an overall aim to improve haematopoietic stem/progenitor cell (HSPC) and mesesnchymal stromal cells (MSC) 3D culture systems, this PhD Thesis addressed the following four interlinked AIMs: (1) The development of a high throughput microwell platform that enabled evaluation of MSC spheroid potential to expand HSPC in vitro; (2) Utilization of the high throughput microwell platform to manufacture HSPC/MSC spheroids to improve the efficacy of direct bone marrow transplantation; (3) The development of an improved microwell platform that retains spheroids within discrete microwells throughout culture; and (4) Characterization of HSPC surface marker change in response to the microwell material.
3
Corner, Helen. "An exploration into transfer of knowledge acquired from taught MSc Human Resource Management (HRM) programmes into workplace Human Resource (HR) Departments and wider dissemination across intra-organisational boundaries." Thesis, University of Derby, 2018. http://hdl.handle.net/10545/622720.
Анотація:
The purpose of this thesis was to explore how knowledge gained during taught Masters in Human Resource Management (MSc HRM) programmes was transferred into working organisations, whether knowledge gained from academic study could be transferred if individuals were motivated to transfer and if organisations had a culture that was receptive to transfer. The term knowledge transfer was defined as sharing of information between one individual and another individual or group. This study looked at the perceived value of Human Resource (HR) knowledge within organisational contexts, with a focus on how knowledge flowed and what facilitated or blocked that flow. A ‘two-tailed’ case study approach was taken using a social construction methodology and was applied across three University Centres, utilising students studying on MSc HRM programmes and their respective work organisations, plus Operational Managers within the same geographical boundaries. Data was gathered using qualitative methods and analysed thematically. A key finding of this study was that knowledge gained from MSc HRM programmes is valued within organisational contexts. HR professionals effectively transferred knowledge into their organisational functions and amongst workplace communities and via wider networks, in a homogenous manner. However, the study also found that transfer of knowledge across work boundaries, via heterogeneous workplace communities, was less effective. Individual willingness to transfer knowledge was found, but issues linked to organisational culture such as politics, power and structure was found to influence the extent of knowledge transfer activities. It was evident that in order for knowledge transfer to be effective an organisational culture based on mutual support and understanding was required. If an organisation had a culture focused on Key Performance Indicators (KPI) that reinforce knowledge transfer across team boundaries then heterogeneous workplace communities emerged. Organisations that deliberately focused on knowledge transfer evidenced a greater ability to transfer knowledge across organisational functions; this strategy was beneficial to organisational growth. This study concluded that building on workplace communities and managing a deliberate introduction of heterogeneous workplace communities enabled MSc HRM acquired-knowledge to be transferred cross organisationally. Although this study focused on the transfer of knowledge from MSc HRM programmes the concept behind using workplace communities to transfer and build knowledge could potentially be transferable to other disciplines. Two further areas of research were identified: firstly, action research within University Centres to ascertain the benefit of cross-discipline teaching, secondly, analysis of an organisation with a heterogeneous community design.
4
Schrön, Felix [Verfasser], Carsten [Akademischer Betreuer] Werner, Carsten [Gutachter] Werner, and Marcy [Gutachter] Zenobi-Wong. "Inkjet bioprinting and 3D culture of human MSC-laden binary starPEG-heparin hydrogels for cartilage tissue engineering / Felix Schrön ; Gutachter: Carsten Werner, Marcy Zenobi-Wong ; Betreuer: Carsten Werner." Dresden : Technische Universität Dresden, 2019. http://d-nb.info/1226944744/34.
5
Monterosso, Melissa Eileen. "The Microwell-mesh platform: A multifaceted microtissue technology to link cell culture, animal models and patients." Thesis, Queensland University of Technology, 2022. https://eprints.qut.edu.au/236375/1/Melissa_Monterosso_Thesis.pdf.
Анотація:
Work towards effective translational medicine has revealed that current in vitro platforms are not sufficient to produce relevant clinical results. Advances in 3D technologies and preclinical models must be made to address the gaps lead to failure of initially promising therapies. The Microwell-mesh platform is a versatile device demonstrated to help bridge this gap. The device within the context of completed work was used to not only generate cancer xenograft models, crucial for successful personalized cancer therapeutics, but was also used to investigate the capacity of adult mesenchymal stem cells to regenerate bone and cartilage tissues within relevant biological environments.
6
Chokder, Rafiul Abedin, and Tapia Paulina Vanessa Díaz. "The role of corporate culture in managing cultural diversity - A case study on a German multinational company." Thesis, Högskolan i Gävle, Avdelningen för ekonomi, 2019. http://urn.kb.se/resolve?urn=urn:nbn:se:hig:diva-29275.
Анотація:
Research Aim: Our aim is to understand how multinational companies integrate cultural diversity of employees in their corporate culture. To achieve this objective, we compare the employees’ perception with the company's view on the topic. Design/methodology/approach: A qualitative case study is conducted with three sets of questionnaires. Two sets of questionnaires were designed for the foreign and the local employees. The third set was created for the department of human resource management who represented the company’s view. Analysis is done by comparing the theories with empirical findings of the study. Findings: The findings revealed that corporate culture is inspired by the national culture. By implementing a proper recruiting process, socialization and teamwork, multinational companies can integrate cultural diversity successfully in their corporate culture. Several tools such as offering language courses, announcements in both languages, a welcoming at the new country booklet, mentors, anonymous feedback on cultural issues and sports or cultural outings are proposed to manage cultural diversity. These tools can be used for both the foreign and the local employees. The integration relies on both employees and the companies. However, upper management should support the department of human resources management to find solutions for the integration of a culturally diverse workforce. Practical implications: Contemporary studies propose tools like mentoring programs that are costly and may ignite stereotyping while managing cultural diversity. This study proposes tools that are cost-effective and functional in integrating and managing cultural diversity of employees. Originality/Value: Previous studies do not emphasize the role of corporate culture in integrating cultural diversity of employees. This study focuses on the empirical gap of employees’ perception on the role of corporate culture in integrating cultural diversity. It proposes, that to manage cultural diversity, companies should only focus on the national and corporate culture of the company and not necessarily of the employee’s culture.Syftet: Vårt mål är att förstå hur multinationella företag integrerar kulturellt mångfald i deras företagskultur. För att uppnå detta mål jämför vi medarbetarnas uppfattning med företagets syn i ämnet. Design / metod / tillvägagångssätt: En kvalitativ fallstudie genomförs med tre uppsättningar av frågeformulär. Två av frågeformulären utformades för utländska och lokala anställda. Den tredje uppsättningen skapades för personalavdelningschefen som representerade företagets uppfattning. En analys görs genom att jämföra teorierna med det empiriska resultatet av studien. Resultat: Resultatet visade att företagskulturen är inspirerad av den nationella kulturen. Genom att implementera en organiserad rekryteringsprocess, socialisering och lagarbete, kan multinationella företagen integrera kulturell mångfald framgångsrikt i sin företagskultur. Flera verktyg så som att erbjuda språkkurser, utskick på bägge språken, ett välkomshäfte för det nya landet, mentorer, anonym feedback om kulturella frågor och sport eller kulturutflykter föreslås för att hantera kulturell mångfald. Dessa verktyg kan användas för både utländska och lokala anställda. Integrationen bygger på både de anställda och företaget. Högre befattningar bör dock stödja personalavdelningen för att hitta lösningar för integration av en multikulturell arbetskraft. Praktiska åtgärder: Samtidsstudier som verktyg så som mentorprogram är kostsamma och kan skapa fördomar samtidigt när man vill behandla ämnet. Den här studien föreslår verktyg som är kostnadseffektiva och funktionella för att integrera och hantera kulturella mångfald hos de anställda. Bidrag: Tidigare studier betonar inte företagskulturens roll i att integrera kulturell mångfald hos anställda. Denna studie fokuserar på det empiriska gapet av medarbetarnas uppfattning om företagskulturens roll för att integrera den kulturella mångfalden. Det föreslås, för att hantera kulturell mångfald bör företagen bara fokusera på den nationella kulturen och företagskulturen och inte nödvändigtvis på medarbetarens kultur.
7
Zomer, Helena Debiazi. "Estabelecimento de cultura de células de pluripotência induzida a partir de células tronco derivadas do tecido adiposo de coelhos." Universidade de São Paulo, 2013. http://www.teses.usp.br/teses/disponiveis/10/10132/tde-18072014-103412/.
Анотація:
As células de pluripotência induzida (iPS) foram reportadas pela primeira vez em 2006 por Takahashi e Yamanaka e desde então vem sendo extensivamente estudadas. Por meio da técnica, células somáticas adultas adquirem comportamento muito semelhante às células tronco embrionárias, reduzindo as questões éticas relacionadas ao uso destas em pesquisas. Entretanto, os mecanismos biológicos das células iPS ainda não estão completamente elucidados. Ainda são necessárias pesquisas mais aprofundadas para garantir a segurança e eficácia de sua possível utilização em futuras terapias. Os coelhos, como modelos experimentais, são vantajosos tanto pelo seu tamanho ideal para procedimentos cirúrgicos quanto por sua manutenção fácil e econômica. As células tronco derivadas do tecido adiposo (ADSC) consistem em um tipo de célula tronco mesenquimal multipotente que se destaca pela facilidade, rapidez e segurança de coleta e processamento. As ADSC foram coletadas e caracterizadas por meio de análises de curva de crescimento celular, doubling time, viabilidade após criopreservação, capacidade de formação de colônias fibroblastoides, diferenciação osteogênica, condrogênica e adipogênica, imunocitoquímica e citometria de fluxo. Elas demonstraram possuir as características básicas de células tronco mesenquimais, destacando-se por uma alta e rápida capacidade proliferativa. A indução de pluripotência foi realizada nas ADSC de coelhos pela introdução de quatro fatores de transcrição (OCT4, SOX2, cMYC, e KLF4), pelo vetor lentiviral STEMCCA (Milipore). Cinco protocolos de indução foram testados. Células resultantes da indução foram caracterizadas quanto à atividade da fosfatase alcalina e perfil fenotípico por citometria de fluxo. A proliferação acentuada das ADSC parece ser um fator limitante para a eficácia da reprogramação. A partir destes dados, espera-se contribuir para o desenvolvimento de uma tecnologia brasileira ainda inédita em coelhos e adicionar informações importantes à literatura, acerca das propriedades das células iPS, visando sua utilização como modelo experimental para futuras terapias.Induced Pluripotent Stem (iPS) cells were first reported in 2006 by Takahashi e Yamanaka and has been intensively studied since then. Through the technique, adult somatic cells acquire behavior very similar to embryonic stem cells, reducing the ethical issues related to the use of such research. However, biological mechanisms of iPS cells are not yet fully elucidated. Thus, further research is needed to ensure the safety and efficacy for their possible use in future therapies. Rabbits, as experimental models, are advantageous by their ideal size for surgical procedures and easy and economic maintenance. Adipose Derived Stem Cells (ADSC) consists of multipotent mesenchymal stem cells that stand for ease, speed and safety of collection and processing. The ADSC were collected and characterized by analysis of cell growth curve, doubling time, viability after cryopreservation, ability of fibroblast colony formation, osteogenic, adipogenic and chondrogenic differentiation, immunocytochemistry and flow cytometry. They showed to have the basic characteristics of mesenchymal stem cells, standing out by a high and fast proliferative capacity. Induction of pluripotency was performed in rabbits ADSC by the introduction of four transcription factors (OCT4, SOX2, cMYC, and KLF4) by lentiviral vector STEMCCA (Millipore). Five protocols were tested and analyzed. The resulting cells after induction were characterized by alkaline phosphatase activity and phenotypic profile by flow cytometry. The great proliferation of ADSC seems to be detrimental for the reprogramming efficiency. From these data, it is expected to contribute to the development of a Brazilian technology still unpublished in rabbits, and to add important information to the literature about the properties of iPS cells, aiming their use as an experimental model for future therapies.
8
Tetenková, Pavla. "Diverzita a interkulturní aspekty fungování MNC." Master's thesis, Vysoká škola ekonomická v Praze, 2015. http://www.nusl.cz/ntk/nusl-261825.
Анотація:
The thesis concentrates on culture, its levels and characteristics, including business culture. The first part focuses on cultural diversity and management of diversity in business culture, with emphasis on the related incorrect tendencies, caused by various culturally conditioned biases. The following part considers different aspects of intercultural communication, its possible barriers and the question of intercultural training. The practical part analyses an existing multinational corporation, particularly with regards to its corporate values, practices, corporate diversity and intercultural training. Furthermore, this part is complemented with a study of the culturally conditioned difficulties within the company, the influence of existing cultural differences on employee communication and cooperation, as well as on intercultural training. The outcome of the study are recommendations, which serve as a base for potential modifications of the corresponding company processes.
9
Inacio, Fatima Pacheco de Santana. "A política de formação de professores em Goiás no contexto dos acordos MEC-USAID (1961-1983)." [s.n.], 2011. http://repositorio.unicamp.br/jspui/handle/REPOSIP/251183.
Анотація:
Orientador: Ediogenes Aragão SantosTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Educação
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Resumo: Este texto analisa a formação de professores, realizada no âmbito das políticas públicas decorrentes dos acordos MEC/USAID/UNESCO, no pós Segunda Guerra Mundial, que resultaram na criação dos Centros de Formação de Professores Primários, entre 1961 e 1983, no município de Catalão, Goiás. Estes cursos de formação, de orientação tecnicista na educação, foram iniciados no Governo Mauro Borges após o Golpe Militar de 1964, com o intuito de planejar, racionalizar os investimentos no setor educacional. Os acordos firmados tinham como elemento chave a formação de uma sociedade "massificada" através da democratização da educação, de bens e de consumo para o mercado. O material didático utilizado nesse processo de formação era elaborado por equipes de trabalho orientadas e vinculadas ao Programa Brasileiro-Americano para o Ensino Primário (PABAEE). As especificidades desta formação, em tempo integral, em regime de internato, se propunha a melhorar a produtividade e a qualidade do ensino do 1º grau e Normal. As principais fontes primárias, inéditas, utilizadas nesta pesquisa foram localizadas no arquivo do Núcleo de Estudos e Pesquisas em Educação de Catalão (NEPEDUCA), da Universidade Federal de Goiás (UFG), Campus Catalão, onde pudemos ter acesso aos projetos para instalação dos Centros Experimentais de Formação de Professores, aos planos de cursos, planilhas, avaliações e textos produzidos pelos professores e bolsistas. A análise das referidas fontes evidenciaram conluio de interesses no período de internacionalização da economia brasileira, criando as condições materiais, técnicas e humanas para a execução dos acordos Brasil/USAID. A interpretação das fontes revelaram-nos que as teorias e metodologias usadas pelos Centro reproduzia uma orientação escolanovista, já ultrapassada nos Estados Unidos, mas que passa a ser usada durante a Ditadura Militar como mecanismo de controle social sem recorrer a violência explícita, pois aplicadas nas escolas, lócus de preservação e distribuição cultural. Os relatos dos bolsistas indicam que mudanças foram sutilmente introduzidas alterando seus valores, comportamentos e práticas pedagógicas, assim como sua percepção de escola, da cidade e das relações socioculturais e políticas engendradas entre as instâncias do poder constituído e a sociedade catalana. As representações construídas pelos bolsistas revelam que os conteúdos transmitidos foram assimilados e condicionaram sua percepção de mundo, de sociedade, e que para o bom funcionamento da cidade havia uma hierarquia entre poderes e posições sociais a ser respeitado, sem contestação e cabia ao indivíduo, agora identificado com os valores da pátria, assumir para si os encargos públicos antes atribuídos ao Estado.
Abstract: This text analyses the teacher training, carried out according to the public policies reached in MEC/USAID/UNESCO agreements, in the post Second World War, which resulted in the creation of Primary Teaching Training Centers, from 1961 to 1983, in Catalão, Goiás. These training courses, with technical orientation on education, started to run during Mauro Borges' Government after the Military Coup in 1964, in order to plan and rationalize the investments in education. The key element of the agreements signed, was the formation of a "mass" society through the democratization of education, of material goods and through the acquisition of things, the market. The material used in this teaching process was developed by teams which were orientated and linked to the Brazilian-American Program for the Primary Education (PABAEE). The specifics of this formation which took place in boarding schools in a full-time period aimed to improve the productivity and the quality of the Elementary School and the "Normal" School. The main primary sources used in this research were found in the files of the "Núcleo de Estudos e Pesquisas em Educação de Catalão (NEPEDUCA) (Study and Research Center for Education, in Catalão), located in the Federal University of Goiás (UFG), Catalão Campus, where it was allowed our access not only to the projects to set up the Experimental Centers for Teaching Training, but also to the lesson plans, worksheets, evaluations and texts created by teachers and scholars with a scholarship. The analysis of the aforementioned sources made it quite clear that during the period of internationalization of the Brazilian economy, the technical, human and material conditions were settled in order to reach Brazil / USAID agreement. The interpretation of the sources showed that the theories and methodologies used by the Centers reproduced a "escolanovista" orientation which wasn't popular in the United States anymore, but which started being used during Military Coup as a social control mechanism without the necessity to make use of explicit violence, as applied in schools, locus of cultural preservation and distribution. The scholars with a scholarship reported that changes were subtly introduced altering values, behaviors and pedagogical practices. Their perception of the school, of the city and of the political and socio-cultural relationships engendered between the power which was constituted and the Catalana society was also altered. The representations built by the scholars with a scholarship show that the contents taught were learnt and understood. Their perception of the world, and the society were conditioned and in order to make the city works and operates well there was an hierarchy of power and social positions to be respected, without any kind of objection, and the individuals, now identified according to the values of the homeland, had to take over public charges previously attributed to the State.
Doutorado
Historia, Filosofia e Educação
Doutor em Educação
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Pina, Fabiana [UNESP]. "O acordo MEC-USAID: ações e reações (1966 – 1968)." Universidade Estadual Paulista (UNESP), 2011. http://hdl.handle.net/11449/93369.
Анотація:
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O presente trabalho procura analisar o Acordo MEC-USAID, buscando destacar sua importância para a estrutura universitária brasileira, as modificações ainda presentes que partiram deste Acordo e o contexto histórico no qual ele foi efetivado. Procuramos desenvolver uma pesquisa que abrangesse três ângulos do Acordo: analisamos o próprio Acordo, fizemos um estudo dos escritores que na época da sua publicação se posicionaram contrários ou favoráveis a ele e, por fim, examinamos a historiografia referente a ele, inclusive dos autores que não o tomaram como tema central
This paper analyzes the MEC-USAID agreement, seeking to highlight its importance to the Brazilian university structure, the changes still present who departed this Agreement and the historical context in which it was accomplished. We seek to develop a survey covering three angles of the Agreement: we analyze the agreement, we made a study of writers at the time of its publication is positioned against or in favor of it and, finally, we examine the historiography related to it, including the authors who not taken as a central theme
Книги з теми "MSC culture":
1
Martins, Maria do Rosa rio Rodrigues., ред. Moc ʹambique, aspectos da cultura material. Coimbra: Universidade de Coimbra, Instituto de Antropologia, 1986.
2
Pangsong, Munhwa. 2010 t'ŭrendŭ weibŭ: MBC k'ŏlch'ŏ rip'ot'ŭ. 8th ed. Sŏul-si: Puk Hausŭ, 2009.
3
LeVitus, Bob. Guide to the Macintosh underground: Mac culture from the inside. Indianapolis, IN: Hayden Books, 1993.
4
Simion, Victor. Mic dicționar de artă sacră și de cultură veche românească. București: Basilica, 2017.
5
Bancks, Tristan. Mac Slater hunts the cool. New York: Simon & Schuster Books for Young Readers, 2010.
6
Bancks, Tristan. Mac Slater vs. the city. New York: Simon & Schuster Books for Young Readers, 2011.
7
Liana, Milanez, ed. Rádio MEC: Herança de um sonho. [Rio de Janeiro, Brazil]: TVE Brasil, 2007.
8
Bancks, Tristan. Mac Slater hunts the cool. New York: Simon & Schuster Books for Young Readers, 2010.
9
Montandon, Mac. Jetpack dreams: One man's up and down (but mostly down) search for the greatest invention that never was. New York: Da Capo Press, 2008.
10
Sanborn, Michelle. Teaching about world cultures: Focus on developing regions. Denver, Colo: Center for Teaching International Relations, University of Denver, 1986.
Частини книг з теми "MSC culture":
1
Vischi, Alessandra. "Employability and Transitions towards Work: MSc Degree Programme in Educational Planning and Human Resource Development, Catholic University of the Sacred Heart of Brescia." In Employability & Competences, 471–80. Florence: Firenze University Press, 2018. http://dx.doi.org/10.36253/978-88-6453-672-9.50.
Анотація:
The acceleration of changes underlines the need to enhance our efforts to adapt education to the dynamics of the current economic situation and the issue of employment. In the framework of the circular economy, pedagogy, which is based on the educability of individuals, takes into consideration forms of educational planning to identify a long-lasting balance between economic prosperity, social wellness, and environmental development. The challenge of the future is the possibility of increasing youth employment; this calls for pedagogical expertise and organizational planning to ensure that everyone’s development is authentic and holistic. To this end, the MSc Degree programme in Educational Planning and Human Resource Development offered by the Catholic University trains graduates to become professional figures with expertise in coordinating and managing the development of human resources (guidance, selection, personal services); the professional training and retraining of project managers in social and educational contexts for both academic and corporate spheres. The guiding vision behind the MSc in Educational Planning and Human Resource Development is fully in line with the Catholic University of the Sacred Heart’s educational project, to support a culture of responsibility and creativity, entrepreneurism and collaboration, multi-disciplinary knowledge and skills, and scientific research for the purpose of holistic human development. Educational planning, in a period of socio-economic and social change, involving the whole planet in many respects, can relaunch an ‘integral model of development’, based on long-term wellbeing, technological innovation, ‘human development’, and the dignity of work
2
Dean, Joan FitzPatrick, and Radvan Markus. "The Internationalist Dramaturgy of Hilton Edwards and Micheál mac Liammóir." In Cultural Convergence, 15–46. Cham: Springer International Publishing, 2020. http://dx.doi.org/10.1007/978-3-030-57562-5_2.
Анотація:
Abstract In part because the Gate explored an experimental dramaturgy, its artistic directors Micheál mac Liammóir and Hilton Edwards often wrote, spoke and advocated for a drama that could move beyond realism. The analysis of Hilton Edwards’s dramatic commentary reaches from his early articles on dramaturgy right up to his encounter with the Berliner Ensemble in 1956 that influenced Edwards’s most elaborate statement on drama, The Mantle of Harlequin (1958). An important part of Edwards’s vision was his cosmopolitanism, his refusal to view drama within a restricted national framework. Nationality, on the other hand, was more important for the self-styled Irishman Micheál mac Liammóir. On close inspection, however, we find that his outlook did not differ much from Edwards’s. Mac Liammóir’s main concern was for Irish(-language) drama to absorb elements from abroad, to escape the straitjacket of Abbey realism and to become distinctive in a global context.
3
Clare, David, and Nicola Morris. "The Transnational Roots of Key Figures from the Early Years of the Gate Theatre, Dublin." In Cultural Convergence, 75–106. Cham: Springer International Publishing, 2020. http://dx.doi.org/10.1007/978-3-030-57562-5_4.
Анотація:
Abstract In Gate Theatre studies, the venue’s original artistic directors, Hilton Edwards and Micheál mac Liammóir, are commonly described as ‘Englishmen’. This chapter breaks new ground by exploring the Irish roots of Edwards and mac Liammóir, and the rumours that mac Liammóir had Spanish and Jewish ancestry. ‘The Boys’ were not the only figures associated with the early Gate to have transnational backgrounds. Coralie Carmichael, the theatre’s biggest female star in its early years, was of mixed Moroccan and Scottish ancestry, and Nancy Beckh, who worked as an actor, costume designer and milliner at the Gate between 1932 and 1956, was a Dubliner of half-German descent. Using critical theories around new interculturalism, the chapter suggests that the mixed backgrounds of these artists helped them to create intercultural performances. It further demonstrates that these performances cannot be simply dismissed as those of people condescendingly engaging in cultural imperialism or shallow cosmopolitanism.
4
Kallunki, Jarmo, Jaakko Kauko, and Oren Pizmony-Levy. "Finland’s Ministry of Education and Culture in the Light of Its Working Groups." In Finland’s Famous Education System, 35–50. Singapore: Springer Nature Singapore, 2023. http://dx.doi.org/10.1007/978-981-19-8241-5_3.
Анотація:
AbstractThe Ministry of Education and Culture (MEC) has traditionally been considered as the most central actor and the powerhouse in the education policy field in Finland. While the position of the MEC in the Finnish education policy system seems stable, there have been several organisational changes within the MEC over the past three decades. One of these is the disintegration of the committee system and its replacement by the working groups system, a trend that is part of a more general change from governing to governance since the 1990s. In this chapter we analyse data containing the MEC’s working groups and their members with social network analysis in order to understand the ways in which the working group system affects the MEC and its operation. Our analysis suggests that the MEC is organised rather strongly by departments: early childhood and general education, vocational education and training, higher education and research, culture and arts, and youth and sports. Analysing the network through the individual working group members we observed that, in addition to public officials, individuals representing interest organisations such as labour and trade unions were important links between the working groups.
5
Sisson, Elaine. "Kismet: Hollywood, Orientalism and the Design Language of Padraic Colum’s Mogu of the Desert." In Cultural Convergence, 175–92. Cham: Springer International Publishing, 2020. http://dx.doi.org/10.1007/978-3-030-57562-5_7.
Анотація:
Abstract The lure of the exotic ‘other’ was implicit from the early years of the Gate’s repertoire. In 1931 the Gate produced Padraic Colum’s Mogu of the Desert, designed by Micheál mac Liammóir and featuring a young Orson Welles. Exploring Mogu uncovers a broader engagement with ‘exotic’ or oriental narratives at the Gate generally. The history and subject matter of Mogu contextualizes mac Liammóir’s fascination with oriental and Middle-Eastern culture within contemporary film. Archival photos illustrate how production stills copied the iconographic styling of film publicity using ‘film-star’ portraiture to promote the Gate. Orientalist narratives require the display of the body through the eroticization of costume – legitimizing the costumed body as a to-be-looked-at space. The Gate’s fascination with oriental settings enables the visibility of ‘transgressive’ sexualities as well as understanding the tastes and appeal of popular culture.
6
Todesco, Gian Marco. "M.C. Escher e il piano iperbolico." In Matematica e cultura 2010, 129–43. Milano: Springer Milan, 2010. http://dx.doi.org/10.1007/978-88-470-1594-4_10.
7
Trabelsi, Khaled, Semy Majoul, Fatma Charfi, and Héla Kallel. "Measles Virus Production in MRC-5 Cells Grown on Microcarriers in a Stirred Bioreactor." In Cells and Culture, 765–69. Dordrecht: Springer Netherlands, 2010. http://dx.doi.org/10.1007/978-90-481-3419-9_133.
8
Pilný, Ondřej. "The Brothers Čapek at the Gate: R.U.R. and The Insect Play." In Cultural Convergence, 141–73. Cham: Springer International Publishing, 2020. http://dx.doi.org/10.1007/978-3-030-57562-5_6.
Анотація:
Abstract The chapter examines two important but hitherto largely neglected productions of famous European dramas at Dublin’s Gate Theatre, R.U.R. (1929, revived 1931) and The Insect Play (1943) by the brothers Čapek, which it attempts to reconstruct insofar as the available documentary evidence allows. In the process, it discusses the complicated textual history of the English versions produced by the Gate and compares their staging by Hilton Edwards and Micheál mac Liammóir with the celebrated original productions in Czechoslovakia. Finally, the reception of the respective productions is juxtaposed, with points of convergence being teased out and the differences brought about by the respective theatrical and political contexts being elucidated.
9
McIvor, Charlotte. "Ghosting Bridgie Cleary: Tom Mac Intyre and Staging This Woman’s Death." In Crossroads: Performance Studies and Irish Culture, 169–79. London: Palgrave Macmillan UK, 2009. http://dx.doi.org/10.1057/9780230244788_14.
10
Ivory, Yvonne. "Prussian Discipline and Lesbian Vulnerability: Christa Winsloe’s Children in Uniform at the Gate." In Cultural Convergence, 193–216. Cham: Springer International Publishing, 2020. http://dx.doi.org/10.1007/978-3-030-57562-5_8.
Анотація:
Abstract This chapter examines the Dublin production and critical reception of Christa Winsloe’s Children in Uniform, which ran to full houses at the Gate for three weeks in April 1934. The play, which deals with the love between a Prussian schoolgirl and her female teacher, had premiered in Leipzig (1930), run successfully in Berlin (1931), and been adapted for the screen as Mädchen in Uniform (1931) before it was translated into English for a successful London run in 1932-1933. Edwards and mac Liammóir probably saw the original German play in Berlin in 1931. Using the prompt copy, lighting plots, photographs and reviews, the chapter shows how Edwards used expressionistic lighting and sonic leitmotifs to underscore the authoritarian regime within which the relationship between the women develops. In following the Berlin staging, Edwards produced a more subversive version of the play than that seen by London audiences or cinema goers.
Тези доповідей конференцій з теми "MSC culture":
1
Kim, Minwook, Jason A. Burdick, and Robert L. Mauck. "Influence of Chondrocyte Zone on Co-Cultures With Mesenchymal Stem Cells in HA Hydrogels for Cartilage Tissue Engineering." In ASME 2012 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2012. http://dx.doi.org/10.1115/sbc2012-80859.
Анотація:
Mesenchymal stem cells (MSCs) are an attractive cell type for cartilage tissue engineering in that they can undergo chondrogenesis in a variety of 3D contexts [1]. Focused efforts in MSC-based cartilage tissue engineering have recently culminated in the formation of biologic materials possessing biochemical and functional mechanical properties that match that of the native tissue [2]. These approaches generally involve the continuous or intermittent application of pro-chondrogenic growth factors during in vitro culture. For example, in one recent study, we showed robust construct maturation in MSC-seeded hyaluronic acid (HA) hydrogels transiently exposed to high levels of TGF-β3 [3]. Despite the promise of this approach, MSCs are a multipotent cell type and retain a predilection towards hypertrophic phenotypic conversion (i.e., bone formation) when removed from a pro-chondrogenic environment (e.g., in vivo implantation). Indeed, even in a chondrogenic environment, many MSC-based cultures express pre-hypertrophic markers, including type X collagen, MMP13, and alkaline phosphatase [4]. To address this issue, recent studies have investigated co-culture of human articular chondrocytes and MSCs in both pellet and hydrogel environments. Chondrocytes appear to enhance the initial efficiency of MSC chondrogenic conversion, as well as limit hypertrophic changes in some instances (potentially via secretion of PTHrP and/or other factors) [5–7]. While these findings are intriguing, articular cartilage has a unique depth-dependent morphology including zonal differences in chondrocyte identity. Ng et al. showed that zonal chondrocytes seeded in a bi-layered agarose hydrogel construct can recreate depth-dependent cellular and mechanical heterogeneity, suggesting that these identities are retained with transfer to 3D culture systems [8]. Further, Cheng et al. showed that differences in matrix accumulation and hypertrophy in zonal chondrocytes was controlled by bone morphogenic protein [9]. To determine whether differences in zonal chondrocyte identity influences MSC fate decisions, we evaluated functional properties and phenotypic stability in photocrosslinked hyaluronic acid (HA) hydrogels using distinct, zonal chondrocyte cell fractions co-cultured with bone marrow derived MSCs.
2
Coughlin, Thomas R., Matthew Haugh, Muriel Voisin, Evelyn Birmingham, Laoise M. McNamara, and Glen L. Niebur. "Primary Cilia Knockdown Reduces the Number of Stromal Cells in Three Dimensional Ex Vivo Culture." In ASME 2013 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2013. http://dx.doi.org/10.1115/sbc2013-14723.
Анотація:
Mesenchymal stem cells (MSCs) are multipotent stromal cells that reside in the bone marrow and differentiate into connective cell lines, such as adipocytes and osteoblasts [1]. An appropriate balance of MSC differentiation toward adipocytes and osteoblasts is vital to bone homeostasis [6]. In vitro work demonstrates that differentiation of MSCs is influenced by mechanical stimuli [2, 3]. In a mouse model, the ratio of adipocytes to MSCs in the marrow was 19% lower compared to controls following treatment by low magnitude mechanical signals (LMMS) [4]. In mice, LMMS increased MSC number by 46% and the differentiation capacity of MSCs was biased towards osteoblastic compared to adipogenic differentiation [5]. Thus, mechanobiological stimuli may play an important role in maintaining balanced MSC differentiation.
3
Huang, Alice H., and Robert L. Mauck. "Extended Long-Term Culture of MSC-Laden Agarose Constructs Does Not Produce Functional Tissue Comparable to Primary Chondrocytes." In ASME 2010 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2010. http://dx.doi.org/10.1115/sbc2010-19643.
Анотація:
Articular cartilage lines the surfaces of joints and transmits the forces arising from locomotion. The poor ability of cartilage to self-repair has motivated efforts to engineer replacements that recapitulate this load-bearing function. While chondrocyte-laden constructs have been generated with near-native mechanical properties, limitations in chondrocyte availability may preclude their clinical use. Therefore, mesenchymal stem cells (MSCs), which can undergo chondrogenesis in 3D culture, have emerged as a promising alternative [1]. However, although MSCs deposit a cartilaginous matrix, mechanical and biochemical properties are lower than those achieved with chondrocytes [1, 2]. Using microarray analysis, we recently showed that limitations in functional MSC chondrogenesis may stem from incomplete or incorrect molecular induction; molecular differences were observed between donor-matched differentiated chondrocytes and newly differentiated MSCs over 8 weeks of culture [2]. While some genes remained consistently low in MSCs compared to chondrocytes, others gradually increased with time, approaching chondrocyte levels by 8 weeks. As these molecules may underlie the functional disparity between chondrocytes and MSCs, we hypothesized that longer culture durations may improve MSC-seeded construct properties and chondrogenesis. To test this hypothesis, we characterized the evolution of functional properties of MSC- and chondrocyte-seeded constructs over 4 months of in vitro culture in pro-chondrogenic medium.
4
Filali, Rayen, Cristian Andrei Badea, Sihem Tebbani, Didier Dumur, Dominique Pareau, and Filipa Lopes. "Interval observer for Chlorella vulgaris culture in a photobioreactor." In Control (MSC). IEEE, 2011. http://dx.doi.org/10.1109/cca.2011.6044419.
5
Farrell, Megan J., Tiffany L. Zachry, and Robert L. Mauck. "Micromechanical Deformation of Chondrogenic Mesenchymal Stem Cells in 3D Hydrogels is Modulated by Time in Culture and Matrix Connectivity." In ASME 2010 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2010. http://dx.doi.org/10.1115/sbc2010-19534.
Анотація:
Mesenchymal stem cells (MSCs) are a clinically attractive alternative to chondrocytes for the development of engineered cartilage tissue owing to their ease of isolation and chondrogenic potential [1]. However, the mechanical properties of MSC-based constructs have yet to match those of native cartilage or of chondrocyte-based constructs cultured similarly [1]. One route for improving these properties may be the application of mechanical stimulation, as normal cartilage development and homeostatic maintenance is dependent on force transduction. In a tissue engineering context, dynamic compression applied to chondrocyte-seeded hydrogels modulates both matrix production and mechanical properties [2, 3]. Similarly, when MSCs are embedded in 3D hydrogels, expression of chondrogenic markers and cartilaginous ECM synthesis are differentially regulated by dynamic compressive loading [4, 5]. Indeed, we have recently shown that long-term dynamic loading initiated after a pre-culture period of 21 days in pro-chondrogenic medium improves matrix distribution and the compressive properties of MSC-seeded constructs [5]. Interestingly, when loading was initiated after a single day of culture, mechanical properties failed to develop [6, 7], suggesting that elaboration of matrix was required prior to dynamic loading in order to positively direct construct maturation. When chondrocytes are embedded in agarose, the initial growth phase is characterized by the establishment of a dense pericellular matrix (PCM). At early times in culture, before these islands of PCM become connected into an interterritorial matrix, cells are protected from bulk deformation applied to the gel [8]. In a recent study, we showed that clonal heterogeneity in stem cell populations determines the rate at which this PCM forms, with some MSC clones rapidly establishing a dense PCM, while others fail to develop a robust PCM (and so continue to deform with gel deformation) through several weeks in culture [9]. To further this investigation, this study charted the culture time-dependent changes in ECM connectivity and MSC deformation under basal and chondrogenic conditions.
6
Erickson, Isaac E., Kilief H. Zellars, Sydney R. Kestle, Jason A. Burdick, and Robert L. Mauck. "Dynamic Compression Promotes Cartilage-Like Functional Properties in MSC-Seeded Hyaluronic Acid Hydrogels." In ASME 2011 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2011. http://dx.doi.org/10.1115/sbc2011-53661.
Анотація:
The specialized function of articular cartilage in distributing stresses during normal joint movement must be recapitulated in a successful engineered cartilage repair. Chondrocytes can generate in vitro cartilage constructs with mechanical properties at or near native levels [1–3] when cultured in specialized media formulations. While these advances in chondrocyte-based tissue engineering are highly instructive, the difficulty of obtaining sufficient numbers of healthy autologous chondrocytes represents a considerable challenge. To circumvent this limitation, many have evaluated mesenchymal stem cells (MSCs), an autologous cell type that can be expanded in vitro and with a demonstrated capacity for chondrogenic differentiation. Despite their potential, MSC-based engineered cartilage has yet to achieve functional properties comparable to those produced by chondrocytes in 3D culture [4–6].
7
Huang, Alice H., and Robert L. Mauck. "Repeated Dynamic Loading Modulates Cartilage Gene Expression but Does Not Improve Mechanical Properties of MSC-Laden Hydrogels." In ASME 2009 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2009. http://dx.doi.org/10.1115/sbc2009-204339.
Анотація:
Mesenchymal stem cells (MSCs) are a multi-potential cell type that can differentiate toward a variety of tissue-specific phenotypes, including cartilage. Given their chondrogenic potential, MSCs are a promising cell source for cartilage tissue engineering (TE). However, while MSCs readily undergo chondrogenesis in 3D culture and deposit a cartilage-like matrix, the mechanical properties of MSC-seeded constructs are greatly inferior to chondrocyte-seeded constructs similarly maintained [1]. To date, optimization strategies for enhancing functional MSC chondrogenesis, including increasing seeding density and transient application of growth factor, have shown limited success [3]. Using microarray analysis, we have recently demonstrated that mis-expression of certain genes, including lubricin, chondromodulin and RGD-CAP, a collagen associated protein, may underlie this disparity in mechanical function [2]. In this study, we examined dynamic compression as an alternative method to enhance MSC differentiation. Previous work using chondrocyte-based constructs have demonstrated that matrix biosynthesis and mechanical properties were improved with the application of cyclic compression [4]. Furthermore, upregulation of lubricin was observed when surface motion was applied to chondrocyte-seeded porous scaffolds [5]. While significant effort has gone toward optimizing loading parameters to direct tissue growth of chondrocyte-based constructs, few studies have examined the effects of mechanical stimulation on MSC-based constructs. Some have demonstrated positive effects on MSC chondrogenesis with application of compressive loading [6, 7], while others have shown that long-term loading may adversely affect the developing mechanical properties of MSC-seeded constructs [8]. In this study, we examined the effects of repeated dynamic compressive loading on MSC chondrogenesis and showed that mechanical properties and gene expression were modulated by this loading modality.
8
Barminko, Jeffrey, Jean Pierre Dolle, Rene Schloss, Martin Grumet, and Martin L. Yarmush. "Encapsulated Mesenchymal Stem Cells for Central Nervous System Repair." In ASME 2010 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2010. http://dx.doi.org/10.1115/sbc2010-19712.
Анотація:
Mesenchymal stromal cells (MSC) have long been regarded as a cell source with the potential to provide therapies for various different tissue pathologies. They were originally identified for their ability to adhere to tissue culture plastic and gained favor due to their tremendous ability to propagate[1]. It was this finding as well as their ability to differentiate into lineages of mesoderm which have long made MSC a potential tool for autologous cellular replacement therapies [2, 3]. More recently, their cyto-protective role has been realized and been implicated in the benefit achieved in treating various different tissue pathologies. MSC have been found to secrete several different cytokines and growth factors in vitro. Furthermore, these factors can be modulated based on the environment MSC are exposed to. MSC have shown therapeutic benefits in models of GVHD, myocardial infarction, fulminant hepatic failure, central nervous system trauma and others, without any apparent cellular replacement. These advances propelled MSC to the fore front of potential cellular therapies and many are seeking to take advantage of their tissue protective properties. However, several draw backs in current methods of MSC implantation limit the ability to carry out safe and controlled clinical trials. Limitation with current MSC implantation approaches include; 1) directly transplanted MSCs exposed to the complex injury environment may be affected themselves early in the treatment processes, 2) MSC may also migrate to undesired tissue locations and 3) may differentiate into undesired end stage cells. These issues severally limit the translatability of MSC treatments in clinical settings; they make controlling experiments very difficult. There becomes a need to develop engineered methods for delivering these cells in a controlled manner. In order to circumvent these potential problems, we propose to use an alginate microencapsulation system as a vehicle for MSC delivery taking advantage of the soluble factors MSC provide.
9
Farrell, Megan J., Eric S. Comeau, and Robert L. Mauck. "Dynamic Culture Improves Mechanical Functionality of MSC-Laden Tissue Engineered Constructs in a Depth-Dependent Manner." In ASME 2011 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2011. http://dx.doi.org/10.1115/sbc2011-53442.
Анотація:
Limitations associated with the use of autologous chondrocytes (CH) for cartilage tissue engineering beget the need for alternative cell sources. Mesenchymal stem cells (MSC) are clinically attractive due to their ability to undergo chondrogenesis in three-dimensional culture [1,2]; however, when compared to CH, MSC fail to develop functional equivalence [2,3]. We have previously shown a marked depth-dependence in local equilibrium modulus of MSC-laden gels, with the superficial zones (where maximal media exchange occurs) considerably stiffer than regions removed from nutrient supply (center and bottom of construct); less dramatic depth-dependence was observed in CH-laden gels [4]. Similarly, other studies have shown depth-dependent properties in CH-laden gels with the construct edge generally stiffer than the center [5]. Given this apparent influence of nutrient supply, the objective of the current study was to assess the impact of dynamic culture (via orbital shaking) on the development of depth-dependent mechanical properties in both MSC and CH-laden hydrogels. Furthermore, we assessed cell viability and matrix content throughout the construct depth to determine the mechanism by which this depth-dependency arises. We hypothesized that improved nutrient transport would reduce construct inhomogeneity (particularly for MSC-laden constructs) and improve bulk mechanical properties.
10
Farrell, Megan J., John Shin, and Robert L. Mauck. "Functional Consequences of Glucose and Oxygen Deprivation in Engineered MSC-Based Cartilage Constructs." In ASME 2013 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2013. http://dx.doi.org/10.1115/sbc2013-14495.
Анотація:
Clinical implementation of stem cell-based cartilage repair techniques has been limited by the inability of these cells to produce cartilaginous tissue equivalent to that produced by native chondrocytes. We have recently shown that while bulk mechanical properties of mesenchymal stem cell (MSC)-laden constructs are lower than chondrocyte-laden constructs, MSCs can in fact produce tissue that matches or exceeds the biochemical and mechanical properties produced by chondrocytes in regions where there is maximal nutrient supply [1]. We also noted that in the central regions of constructs, where nutrient and oxygen availability is lowest (due to consumption through the construct depth), MSC viability was markedly lower than in the outer regions and drastically lower than the center of chondrocyte-laden constructs maintained similarly. These data suggest that MSCs can achieve a high anabolic functionality when they undergo chondrogenesis (via the provision of TGF-β3) and in doing so can produce tissue of equivalent or greater properties than chondrocytes. However, unlike chondrocytes, MSCs appear thrive only when they are provided with a sufficient nutrient supply. To further delineate the role of microenvironmental stressors [2, 3, 4] on MSC viability and functional capacity, we evaluated the impact of glucose and oxygen deprivation, in the presence and absence of TGF-β, during long term culture. Furthermore, since MSC isolation procedures result in a heterogeneous cell population [5,6], we investigated whether different clonal populations respond to these microenvironmental stressors in a distinct fashion.
Звіти організацій з теми "MSC culture":
1
Wick, Charles H., and Patrick E. McCubbin. Removing Complex Growth Media from MS2 Bacteriophage Cultures. Fort Belvoir, VA: Defense Technical Information Center, August 1999. http://dx.doi.org/10.21236/ada368537.
2
Jact. L52122 Mitigation of MIC using CP Under Disbonded Coating. Chantilly, Virginia: Pipeline Research Council International, Inc. (PRCI), January 2008. http://dx.doi.org/10.55274/r0011099.
Анотація:
This report summarizes lab work carried out in a test cell in which line pipe steel was exposed under anaerobic conditions to a freshly prepared synthetic corrosion product consisting of iron sulfide and iron carbonate under a permeable �coating� barrier. Weight loss for the steel specimen was sharply reduced by application of CP with rates of corrosion for short term experiments dropping to less than 0.05 mm/year at potentials more negative than �950 mVCSE at the pipe surface. This value is in good agreement with past literature. Addition of a mixed microbial culture including SRB to the experiment gave similar results in terms of weight loss.
3
Pound. L52104 Differentiation of Corrosion Mechanisms by Morphological Feature Characterization. Chantilly, Virginia: Pipeline Research Council International, Inc. (PRCI), August 2004. http://dx.doi.org/10.55274/r0011097.
Анотація:
Corrosion of liquid and gas pipelines can occur by various mechanisms. The ability to differentiate between mechanisms is crucial if corrosion control measures are to be effective. The objective of this work was to determine whether corrosion of pipeline steels results in characteristic morphological features that are diagnostic for specific corrosion mechanisms, particularly with regard to microbiologically influenced corrosion (MIC). Coupons of 1018 carbon steel were exposed for two weeks in 5 wt% NaCl under abiotic and biotic conditions in different environments (N2, N2-CO2, and N2-H2S). Pitting occurred in all environments both with and without bacteria present. Many of the pits formed under biotic conditions were similar in morphology to those formed under abiotic conditions. However, other pits exhibited a different morphology from the abiotic pits in the N2 and N2-CO2 environments. In the N2-H2S environment, the presence of bacteria did not result in any discernible differences in pit morphology. The biotic pits in the N2 and N2-CO2 environments were similar in shape and size to those previously found on pipeline steel in a biotic culture medium, where MIC was essentially the sole cause of pitting. Thus, identification of pits associated with MIC appears feasible for natural gas environments.
4
Romero Ramírez, Álvaro Fernando, and Felipe Eugenio Mora Parra. Plan estratégico y prospectivo del municipio de la Plata Huila como eje regional de desarrollo de turismo sostenible 2020 – 2030. Universidad Nacional Abierta y a Distancia, 2021. http://dx.doi.org/10.22490/ecacen.5439.
Анотація:
El turismo en Colombia, es un sector importante en la economía; el departamento del Huila posee un potencial alto de desarrollo en este sector; este trabajo se realizó en el municipio de La Plata, en un ámbito de desarrollo turístico sostenible; entendiendo esto como el equilibrio entre el sector privado, publico, académico y la comunidad. El objetivo del presente trabajo fue estructurar el plan estratégico y prospectivo del municipio de La Plata Huila para convertirlo como eje regional de desarrollo en el turismo sostenible 2020 – 2030. Para esto se utilizaron los métodos de prospectiva estratégica planteados por Mojica (2010); se desarrolló el análisis del sector, a través de estudios bibliométricos, patentometría, vigilancia tecnológica, aplicaciones como Mic-Mac, Mactor y ábaco de Regniere. También se realizó la estructuración de variables estratégicas, el análisis morfológico de los escenarios, la postulación de estos escenarios en los ejes de Peter Schwartz y la determinación del grado de gobernabilidad de las acciones a desarrollar. Los resultados mostraron que el desarrollo del sector turístico en el mundo, ha tenido como eje fundamental el turismo sostenible integrado con el ámbito rural. El escenario “La Plata Ciudad Región” fue elegido como reto, y en este se aprovecharon los potenciales del turismo rural y cultural, permitiendo que los turistas hicieran una inmersión en las actividades y los ambientes tradicionales. Además, presenta alianzas entre el estado y la empresa privada para potencializar estos cambios; en este escenario el turista disfruta de un parque automotor acondicionado, pensando en la comodidad y la seguridad.
5
Heifetz, Yael, and Michael Bender. Success and failure in insect fertilization and reproduction - the role of the female accessory glands. United States Department of Agriculture, December 2006. http://dx.doi.org/10.32747/2006.7695586.bard.
Анотація:
The research problem. Understanding of insect reproduction has been critical to the design of insect pest control strategies including disruptions of mate-finding, courtship and sperm transfer by male insects. It is well known that males transfer proteins to females during mating that profoundly affect female reproductive physiology, but little is known about the molecular basis of female mating response and no attempts have yet been made to interfere with female post-mating responses that directly bear on the efficacy of fertilization. The female reproductive tract provides a crucial environment for the events of fertilization yet thus far those events and the role of the female tract in influencing them are poorly understood. For this project, we have chosen to focus on the lower reproductive tract because it is the site of two processes critical to reproduction: sperm management (storage, maintenance, and release from storage) and fertilization. E,fforts during this project period centered on the elucidation of mating responses in the female lower reproductive tract The central goals of this project were: 1. To identify mating-responsive genes in the female lower reproductive tract using DNA microarray technology. 2. In parallel, to identify mating-responsive genes in these tissues using proteomic assays (2D gels and LC-MS/MS techniques). 3. To integrate proteomic and genomic analyses of reproductive tract gene expression to identify significant genes for functional analysis. Our main achievements were: 1. Identification of mating-responsive genes in the female lower reproductive tract. We identified 539 mating-responsive genes using genomic and proteomic approaches. This analysis revealed a shift from gene silencing to gene activation soon after mating and a peak in differential gene expression at 6 hours post-mating. In addition, comparison of the two datasets revealed an expression pattern consistent with the model that important reproductive proteins are pre-programmed for synthesis prior to mating. This work was published in Mack et al. (2006). Validation experiments using real-time PCR techniques suggest that microarray assays provide a conservativestimate of the true transcriptional activity in reproductive tissues. 2.lntegration of proteomics and genomics data sets. We compared the expression profiles from DNA microarray data with the proteins identified in our proteomic experiments. Although comparing the two data sets poses analyical challenges, it provides a more complete view of gene expression as well as insights into how specific genes may be regulated. This work was published in Mack et al. (2006). 3. Development of primary reproductive tract cell cultures. We developed primary cell cultures of dispersed reproductive tract cell types and determined conditions for organ culture of the entire reproductive tract. This work will allow us to rapidly screen mating-responsive genes for a variety of reproductive-tract specifi c functions. Scientific and agricultural significance. Together, these studies have defined the genetic response to mating in a part of the female reproductive tract that is critical for successful fertllization and have identified alarge set of mating-responsive genes. This work is the first to combine both genomic and proteomic approaches in determining female mating response in these tissues and has provided important insights into insect reproductive behavior.
6
Jorgensen, Frieda, John Rodgers, Daisy Duncan, Joanna Lawes, Charles Byrne, and Craig Swift. Levels and trends of antimicrobial resistance in Campylobacter spp. from chicken in the UK. Food Standards Agency, September 2022. http://dx.doi.org/10.46756/sci.fsa.dud728.
Анотація:
Campylobacter spp. are the most common bacterial cause of foodborne illness in the UK, with chicken considered to be the most important vehicle of transmission for this organism. It is estimated there are 500,000 cases of campylobacteriosis in the UK annually, with Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli) accounting for approximately 91% and 8 % of infections, respectively. Although severe infection in humans is uncommon, treatment is seldom needed for human infection but usually involves the administration of a macrolide (e.g., azithromycin) or a fluoroquinolone (e.g., ciprofloxacin). An increased rate of resistance in Campylobacter in chicken to such antimicrobials could limit effective treatment options for human infections and it is therefore important to monitor changes in rates of resistance over time. In this report we analysed trends in antimicrobial resistance (AMR) in C. jejuni and C. coli isolated from chicken in the UK. The chicken samples were from chicken reared for meat (ie. broiler chicken as opposed to layer chicken (ie. egg-laying chicken)) and included chicken sampled at slaughterhouses as well as from retail stores in the UK. Datasets included AMR results from retail surveys of Campylobacter spp. on chicken sampled in the UK from various projects in the time period from 2001 to 2020. In the retail surveys, samples were obtained from stores including major and minor retail stores throughout the UK (in proportion to the population size of each nation) and Campylobacter spp. testing was performed using standard methods with the majority of isolates obtained from direct culture on standard media (mCCDA). Data from national scale surveys of broiler chicken, sampling caecal contents and carcase neckskins at slaughterhouses, undertaken by APHA in 2007/2008, and between 2012 and 2018 were also included in the study. In the APHA-led surveys, Campylobacter were isolated using standard culture methods (culture onto mCCDA) and antimicrobial susceptibility testing was performed by a standard microbroth dilution method to determine the minimum inhibitory concentration (MIC) of isolates. Care was taken when comparing data from different studies as there had been changes to the threshold used to determine if an isolate was susceptible or resistant to an antimicrobial in a small number of scenarios. Harmonised thresholds (using epidemiological cut-off (ECOFF) values) were employed to assess AMR with appropriate adjustments made where required to allow meaningful comparisons of resistance prevalence over time. Data from additional isolates where resistance to antimicrobials were predicted from genome sequence data were also considered.
7
Levisohn, Sharon, Maricarmen Garcia, David Yogev, and Stanley Kleven. Targeted Molecular Typing of Pathogenic Avian Mycoplasmas. United States Department of Agriculture, January 2006. http://dx.doi.org/10.32747/2006.7695853.bard.
Анотація:
Intraspecies identification (DNA "fingerprinting") of pathogenic avian mycoplasmas is a powerful tool for epidemiological studies and monitoring strain identity. However the only widely method available for Mycoplasma gallisepticum (MG) and M. synoviae (MS)wasrandom amplified polymorphic DNA (RAPD). This project aimed to develop alternative and supplementary typing methods that will overcome the major constraints of RAPD, such as the need for isolation of the organism in pure culture and the lack of reproducibility intrinsic in the method. Our strategy focussed on recognition of molecular markers enabling identification of MG and MS vaccine strains and, by extension, pathogenic potential of field isolates. Our first aim was to develop PCR-based systems which will allow amplification of specific targeted genes directly from clinical material. For this purpose we evaluated the degree of intraspecies heterogeneity in genes encoding variable surface antigens uniquely found in MG all of which are putative pathogenicity factors. Phylogenic analysis of targeted sequences of selected genes (pvpA, gapA, mgc2, and lp) was employed to determine the relationship among MG strains.. This method, designated gene targeted sequencing (GTS), was successfully employed to identify strains and to establish epidemiologically-linked strain clusters. Diagnostic PCR tests were designed and validated for each of the target genes, allowing amplification of specific nucleotide sequences from clinical samples. An mgc2-PCR-RFLP test was designed for rapid differential diagnosis of MG vaccine strains in Israel. Addressing other project goals, we used transposon mutagenesis and in vivo and in vitro models for pathogenicity to correlated specific changes in target genes with biological properties that may impact the course of infection. An innovative method for specific detection and typing of MS strains was based on the hemagglutinin-encoding gene vlhA, uniquely found in this species. In parallel, we evaluated the application of amplified fragment length polymorphism (AFLP) in avian mycoplasmas. AFLP is a highly discriminatory method that scans the entire genome using infrequent restriction site PCR. As a first step the method was found to be highly correlated with other DNA typing methods for MG species and strain differentiation. The method is highly reproducible and relatively rapid, although it is necessary to isolate the strain to be tested. Both AFLP and GTS are readily to amenable to computer-assisted analysis of similarity and construction of a data-base resource. The availability of improved and diverse tools will help realize the full potential of molecular typing of avian mycoplasmas as an integral and essential part of mycoplasma control programs.
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Borch, Thomas, Yitzhak Hadar, and Tamara Polubesova. Environmental fate of antiepileptic drugs and their metabolites: Biodegradation, complexation, and photodegradation. United States Department of Agriculture, January 2012. http://dx.doi.org/10.32747/2012.7597927.bard.
Анотація:
Many pharmaceutical compounds are active at very low doses, and a portion of them regularly enters municipal sewage systems and wastewater-treatment plants following use, where they often do not fully degrade. Two such compounds, CBZ and LTG, have been detected in wastewater effluents, surface waters, drinking water, and irrigation water, where they pose a risk to the environment and the food supply. These compounds are expected to interact with organic matter in the environment, but little is known about the effect of such interactions on their environmental fate and transport. The original objectives of our research, as defined in the approved proposal, were to: Determine the rates, mechanisms and products of photodegradation of LTG, CBZ and selected metabolites in waters exposed to near UV light, and the influence of DOM type and binding processes on photodegradation. Determine the potential and pathways for biodegradation of LTG, CBZ and selected metabolites using a white rot fungus (Pleurotusostreatus) and ADP, and reveal the effect of DOM complexation on these processes. Reveal the major mechanisms of binding of LTG, CBZ and selected metabolites to DOM and soil in the presence of DOM, and evaluate the effect of this binding on their photodegradation and/or biodegradation. We determined that LTG undergoes relatively slow photodegradation when exposed to UV light, and that pH affects each of LTG’s ability to absorb UV light, the efficiency of the resulting reaction, and the identities of LTG’sphotoproducts (t½ = 230 to 500 h during summer at latitude 40 °N). We observed that LTG’sphotodegradation is enhanced in the presence of DOM, and hypothesized that LTG undergoes direct reactions with DOM components through nucleophilic substitution reactions. In combination, these data suggest that LTG’s fate and transport in surface waters are controlled by environmental conditions that vary with time and location, potentially affecting the environment and irrigation waters. We determined that P. ostreatusgrows faster in a rich liquid medium (glucose peptone) than on a natural lignocellulosic substrate (cotton stalks) under SSF conditions, but that the overall CBZ removal rate was similar in both media. Different and more varied transformation products formed in the solid state culture, and we hypothesized that CBZ degradation would proceed further when P. ostreatusand the ᵉⁿᶻʸᵐᵃᵗⁱᶜ ᵖʳᵒᶠⁱˡᵉ ʷᵉʳᵉ ᵗᵘⁿᵉᵈ ᵗᵒ ˡⁱᵍⁿⁱⁿ ᵈᵉᵍʳᵃᵈᵃᵗⁱᵒⁿ. ᵂᵉ ᵒᵇˢᵉʳᵛᵉᵈ ¹⁴C⁻Cᴼ2 ʳᵉˡᵉᵃˢᵉ ʷʰᵉⁿ ¹⁴C⁻ᶜᵃʳᵇᵒⁿʸˡ⁻ labeled CBZ was used as the substrate in the solid state culture (17.4% of the initial radioactivity after 63 days of incubation), but could not conclude that mineralization had occurred. In comparison, we determined that LTG does not degrade in agricultural soils irrigated with treated wastewater, but that P. ostreatusremoves up to 70% of LTG in a glucose peptone medium. We detected various metabolites, including N-oxides and glycosides, but are still working to determine the degradation pathway. In combination, these data suggest that P. ostreatuscould be an innovative and effective tool for CBZ and LTG remediation in the environment and in wastewater used for irrigation. In batch experiments, we determined that the sorption of LTG, CBZ and selected metabolites to agricultural soils was governed mainly by SOM levels. In lysimeter experiments, we also observed LTG and CBZ accumulation in top soil layers enriched with organic matter. However, we detected CBZ and one of its metabolites in rain-fed wheat previously irrigated with treated wastewater, suggesting that their sorption was reversible, and indicating the potential for plant uptake and leaching. Finally, we used macroscale analyses (including adsorption/desorption trials and resin-based separations) with molecular- level characterization by FT-ICR MS to demonstrate the adsorptive fractionation of DOM from composted biosolids by mineral soil. This suggests that changes in soil and organic matter types will influence the extent of LTG and CBZ sorption to agricultural soils, as well as the potential for plant uptake and leaching.
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Porat, Ron, Doron Holland, and Linda Walling. Identification of Citrus Fruit-Specific and Pathogen-Induced Promoters and Their Use in Molecular Engineering. United States Department of Agriculture, January 2001. http://dx.doi.org/10.32747/2001.7585202.bard.
Анотація:
This one year BARD project was funded to develop methods to monitor promoter activity a gene expression patterns in citrus fruit. To fulfill this goal, we divided the research tasks between both labs so that the Israeli side evaluated the use of microprojectile bombardment ; a tool to evaluate transient gene expression in various citrus fruit tissues, and the US side optimized technical parameters required for Agrobacterium-mediated transformation of various citrus cultivars. Microprojectile bombardment appeared to be a very efficient method for transient gene expression analysis in citrus leaf tissues but was somewhat less applicable in fruit tissues. Nevertheless, we did succeeded to achieve significant levels of 35S-GUS gene expression in young green flavedo tissue. However, only single random spots of 35S-GUS gene expression were detected mature flavedo and in juice sacs and albedo tissue. Overall, we assume that following some more technical improvements particle bombardment could provide a useful technique to rapidly analyze promoter activity at least in the flavedo tissue. For Agrobacterium-mediated transformation, we found that shoot cultures of 'Washington' navel oranges,'Fairchild' mandarins,'Eureca' lemons,'Troyer' citrange and various grapefruits provided a more reliable and consistent source of tissue for transformation than germinated seedlings. Moreover, various growth media's (McCown, Quoirin & Lepoivre, DCR) further improved shoot and root growth relative to MS mineral media, which is commonly used. Also pure white light (using bulbs which do not emit UV or blue light) improved shoot growth in various citrus varieties, and paromomycin appeared to be a more efficient antibiotic for the selection of transgenic plants than Kanamycin. Overall, these optimizations improve transformation efficacy and shoot growth and rooting capacity. In addition to the development of transformation methods, both Israeli and US labs achieved progress in the identification of citrus fruit-specific promoters. In Israel, we isolated a 3.6 kb promoter fragment of the thiamine biosynthesis c-thi gene, which is highly expressed in fruit peel tissue, whereas in the US we isolated a 1.5 kb promoter fragment of the citrus seed-specific cDNA CssH. The identification of more fruit-specific cDNAs and their corresponding promoter regions is currently in progress.
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‘Understanding developmental cognitive science from different cultural perspectives’ – In Conversation with Tochukwu Nweze. ACAMH, October 2020. http://dx.doi.org/10.13056/acamh.13666.
Анотація:
Tochukwu Nweze, lecturer in the Department of Psychology, University of Nigeria, Nsukka and, PhD student in MRC Cognition and Brain Sciences Unit, University of Cambridge talks about his recent paper on parentally deprived Nigerian children having enhanced working memory ability, how important is it to study cultural differences in cognitive adaption during and following periods of adversity, and how can mental health professionals translate this understanding of difference into their work.