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Tsuji, K., T. Nakahata, M. Takagi, T. Kobayashi, A. Ishiguro, T. Kikuchi, K. Naganuma, K. Koike, A. Miyajima, and K. Arai. "Effects of interleukin-3 and interleukin-4 on the development of "connective tissue-type" mast cells: interleukin-3 supports their survival and interleukin-4 triggers and supports their proliferation synergistically with interleukin-3." Blood 75, no. 2 (January 15, 1990): 421–27. http://dx.doi.org/10.1182/blood.v75.2.421.421.
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Abstract We examined the effects of interleukin-3 (IL-3) and interleukin-4 (IL- 4) on connective tissue-type mast cells (CTMC) purified from murine peritoneal cells. Although both factors failed to induce extensive proliferation of CTMC, they stimulated CTMC proliferation synergistically in a dose-dependent manner. Pretreatment of CTMC with IL-3 and/or IL-4 indicated that the sustained presence of both factors was required for the development of type 1 mast cell colonies. The delayed addition of IL-3 to cultures of purified CTMC with IL-4 induced no colony formation, while the delayed addition of IL-4 to cultures with IL-3, even on day 28 of culture, induced type 1 colony formation. In replating type 1 colonies induced by IL-3 and IL-4 to secondary cultures with IL-3 alone, few secondary colonies developed. However, the delayed addition of IL-4 to the secondary culture induced many type 1 colonies. The purified CTMC cultured with IL-3 retained the morphological and cytochemical characteristics of CTMC, as well as proliferative ability. These observations indicate that IL-3 supports the survival of CTMC in methylcellulose culture and that IL-4 triggers and supports CTMC proliferation synergistically with IL-3. The serum- free culture of purified CTMC and the culture of single CTMC demonstrated that the synergistic effect of IL-3 and IL-4 on colony growth and the surviving effect of IL-3 on CTMC require no influence from accessory cells or other humoral factors.
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Tsuji, K., T. Nakahata, M. Takagi, T. Kobayashi, A. Ishiguro, T. Kikuchi, K. Naganuma, K. Koike, A. Miyajima, and K. Arai. "Effects of interleukin-3 and interleukin-4 on the development of "connective tissue-type" mast cells: interleukin-3 supports their survival and interleukin-4 triggers and supports their proliferation synergistically with interleukin-3." Blood 75, no. 2 (January 15, 1990): 421–27. http://dx.doi.org/10.1182/blood.v75.2.421.bloodjournal752421.
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We examined the effects of interleukin-3 (IL-3) and interleukin-4 (IL- 4) on connective tissue-type mast cells (CTMC) purified from murine peritoneal cells. Although both factors failed to induce extensive proliferation of CTMC, they stimulated CTMC proliferation synergistically in a dose-dependent manner. Pretreatment of CTMC with IL-3 and/or IL-4 indicated that the sustained presence of both factors was required for the development of type 1 mast cell colonies. The delayed addition of IL-3 to cultures of purified CTMC with IL-4 induced no colony formation, while the delayed addition of IL-4 to cultures with IL-3, even on day 28 of culture, induced type 1 colony formation. In replating type 1 colonies induced by IL-3 and IL-4 to secondary cultures with IL-3 alone, few secondary colonies developed. However, the delayed addition of IL-4 to the secondary culture induced many type 1 colonies. The purified CTMC cultured with IL-3 retained the morphological and cytochemical characteristics of CTMC, as well as proliferative ability. These observations indicate that IL-3 supports the survival of CTMC in methylcellulose culture and that IL-4 triggers and supports CTMC proliferation synergistically with IL-3. The serum- free culture of purified CTMC and the culture of single CTMC demonstrated that the synergistic effect of IL-3 and IL-4 on colony growth and the surviving effect of IL-3 on CTMC require no influence from accessory cells or other humoral factors.
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Miller, G. "ANIMAL BEHAVIOR: Tool Study Supports Chimp Culture." Science 309, no. 5739 (August 26, 2005): 1311a. http://dx.doi.org/10.1126/science.309.5739.1311a.
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Neathery, Melissa. "Creating a Culture that Supports Mental Health." Journal of Christian Nursing 41, no. 1 (January 2024): 10. http://dx.doi.org/10.1097/cnj.0000000000001132.
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Schumacher, Anne, Eileen Poloski, and Ana Zenclussen. "Human chorionic gonadotropin supports human pregnancy by induction of regulatory T cells (MUC7P.766)." Journal of Immunology 192, no. 1_Supplement (May 1, 2014): 197.18. http://dx.doi.org/10.4049/jimmunol.192.supp.197.18.
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Abstract Regulatory T cells (Treg) are proposed to be involved in fetal tolerance induction. However, the precise mechanisms underlying their generation and function are still not defined. Recently, we proved that the pregnancy hormone human Chorionic Gonadotropin (hCG) efficiently attracts human Treg to trophoblasts favoring a local Treg accumulation. Here we analyzed whether hCG is also involved in Treg generation and activity. Naïve T cells were isolated from blood of pregnant women and co-cultured with either hCG-producing trophoblasts (JEG-3, primary cells) or non-hCG-producing trophoblasts (HTR8, Swan71) and keratinocytes (HaCat). To confirm hCG specificity, we either blocked hCG in JEG-3 co-cultures or used urine-purified or recombinant hCG as culture supplements. Treg induction was then assessed by the expression of specific Treg markers. Furthermore suppressive function of hCG-treated naïve T cells was evaluated in a mixed leukocyte reaction. Co-culture of naïve T cells with JEG-3, primary trophoblasts or supplemented hCG resulted in a statistically significant increase in the expression of specific Treg markers as compared to co-cultures with HTR8, Swan71 or HaCat cells. Moreover, blockage of hCG in JEG-3 co-cultures impaired the up-regulation of Treg markers. Interestingly, we further confirmed an increased suppressive activity of naïve T cells after hCG treatment. Our results strongly suggest a key role for hCG in Treg generation and induction of suppressive activity.
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Wen, Bo, Hui Li, Daru Lu, Xiufeng Song, Feng Zhang, Yungang He, Feng Li, et al. "Genetic evidence supports demic diffusion of Han culture." Nature 431, no. 7006 (September 2004): 302–5. http://dx.doi.org/10.1038/nature02878.
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Morales-Román, Rosa M., Maria Guillot-Ferriols, Laura Roig-Pérez, Senentxu Lanceros-Mendez, Gloria Gallego-Ferrer, and José Luis Gómez Ribelles. "Freeze-extraction microporous electroactive supports for cell culture." European Polymer Journal 119 (October 2019): 531–40. http://dx.doi.org/10.1016/j.eurpolymj.2019.07.011.
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Pawlak, Kim. "Build a Culture That Supports Work‐Life Balance." Nonprofit Communications Report 21, no. 9 (August 10, 2023): 2. http://dx.doi.org/10.1002/npcr.32236.
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Sassi, Lisa, Omolola Ajayi, Sara Campinoti, Dipa Natarajan, Claire McQuitty, Riccardo Rayan Siena, Sara Mantero, et al. "A Perfusion Bioreactor for Longitudinal Monitoring of Bioengineered Liver Constructs." Nanomaterials 11, no. 2 (January 21, 2021): 275. http://dx.doi.org/10.3390/nano11020275.
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In the field of in vitro liver disease models, decellularised organ scaffolds maintain the original biomechanical and biological properties of the extracellular matrix and are established supports for in vitro cell culture. However, tissue engineering approaches based on whole organ decellularized scaffolds are hampered by the scarcity of appropriate bioreactors that provide controlled 3D culture conditions. Novel specific bioreactors are needed to support long-term culture of bioengineered constructs allowing non-invasive longitudinal monitoring. Here, we designed and validated a specific bioreactor for long-term 3D culture of whole liver constructs. Whole liver scaffolds were generated by perfusion decellularisation of rat livers. Scaffolds were seeded with Luc+HepG2 and primary human hepatocytes and cultured in static or dynamic conditions using the custom-made bioreactor. The bioreactor included a syringe pump, for continuous unidirectional flow, and a circuit built to allow non-invasive monitoring of culture parameters and media sampling. The bioreactor allowed non-invasive analysis of cell viability, distribution, and function of Luc+HepG2-bioengineered livers cultured for up to 11 days. Constructs cultured in dynamic conditions in the bioreactor showed significantly higher cell viability, measured with bioluminescence, distribution, and functionality (determined by albumin production and expression of CYP enzymes) in comparison to static culture conditions. Finally, our bioreactor supports primary human hepatocyte viability and function for up to 30 days, when seeded in the whole liver scaffolds. Overall, our novel bioreactor is capable of supporting cell survival and metabolism and is suitable for liver tissue engineering for the development of 3D liver disease models.
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Fujishiro, Aya, Yasuo Miura, Masaki Iwasa, Sumie Fujii, Akihiro Tamura, Atsushi Sato, Asumi Yokota, et al. "Vitamin K2 Supports Hematopoiesis through Acting on Bone Marrow Mesenchymal Stromal/Stem Cells." Blood 126, no. 23 (December 3, 2015): 1192. http://dx.doi.org/10.1182/blood.v126.23.1192.1192.
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Abstract [Background] Myelodysplastic syndrome is an intractable disorder characterized by ineffective hematopoiesis. Although allogeneic hematopoietic stem cell transplantation is the only curative therapy for eligible patients, hematopoiesis-supportive pharmacotherapy is practically important for transplant-ineligible patients to overcome transfusion dependency and infections. Vitamin K2 (VK2, menatetrenone) is a drug used to aim at improvement of hematopoiesis in MDS patients (Leukemia 14: 1156, 2000). However, the exact mechanism how VK2 improves hematopoiesis remains largely unknown. It was reported that VK2 induces MDS cells to undergo apoptosis (Leukemia 13: 1399, 1999). Here, we investigated our hypothesis that VK2 exerts its hematopoiesis-supportive effects through acting on mesenchymal stem/stromal cells (BM-MSCs) in the bone marrow microenvironment. [Methods] Normal bone marrow (BM) samples from healthy adult volunteers were purchased from AllCells (Emeryville, CA). BM-CD34+ cells were isolated from BM-mononuclear cells using anti-CD34 immunomagnetic microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany). Human BM-MSCs were isolated according to our previously published methods (Stem Cells 32:2245, 2014). In co-culture experiments, BM-MSCs with or without VK2 treatment were seeded on a 24-well culture plate. BM-CD34+ cells were applied on the MSC-grown plate and co-cultured in SFEM (StemCell Technologies, Vancouver, Canada) supplemented with 100 ng/mL SCF, 100 ng/mL Flt-3 ligand, 50 ng/mL TPO and 20 ng/mL IL-3. After 10 days of co-culture, the number and surface marker expression of the expanded hematopoietic cells were examined by flow cytometric analysis. [Results] We first tested the direct effect of VK2 on BM-CD34+ cells. BM-CD34+ cells were treated with VK2 at various concentrations ranged from 0 µM to 10 µM for 24 hours and then cultured in SFEM in combinations with cytokines. Surprisingly, viable hematopoietic cells were hardly detected in the expansion culture of BM-CD34+ cells treated with 10 µM VK2. Even with 1 µM treatment, the number of CD45+ cells was decreased, as compared to that of expansion culture of untreated BM-CD34+ cells. The apoptosis analysis showed that the percentage of AnnexinV+ PI+ cells in the expanded hematopoietic cells is increased by VK2 treatment. We next examined the effect of VK2 on the hematopoiesis-supportive capability of BM-MSCs. BM-MSCs were pretreated with VK2 at various concentrations and then co-cultured with BM-CD34+ cells. The numbers of CD34+ cells and CD45+ cells were increased in a VK2 dose-dependent manner. These results demonstrated that VK2 shows different effects on distinct stem/progenitor cells: the induction of apoptosis in BM-CD34+ cells and the enhancement of hematopoiesis-supportive capability of BM-MSCs. We then investigated whether apoptosis-related cell death of BM-CD34+ cells by VK2 treatment is ameliorated in the presence of BM-MSCs. Both BM-CD34+ cells and BM-MSCs were treated with VK2 for 24 hours, and then co-cultured. The number of CD34+ cells was not decreased significantly in contrast to its severe decrease in single culture of VK2-treated BM-CD34+ cells. We further analyzed the effect of VK2 on BM-MSCs. Subpopulation analysis in co-culture of CD34+ cells with VK2-treated BM-MSCs showed that the expansion efficacy of CD34+CD38+ cells is higher in comparison to that of CD34+CD38- cells. In addition, the percentages of CD34-CD33+ cells and CD34-CD13+ cells were higher than those in co-cultures with untreated BM-MSCs. Therefore, VK2-treated BM-MSCs supported the expanded CD34+ cells to skew their phenotype toward myeloid lineage. The presence of a transwell in the co-culture system was unrelated to the expansion pattern of CD34+ cells, which suggested the involvement of soluble factors with respect to the underlining mechanism. We therefore compared the levels of hematopoiesis-supporting cytokine mRNA expression in VK2-treated and untreated BM-MSCs: VK2-treated BM-MSCs showed lower expression of CXCL12/SDF-1 mRNA and a trend toward higher expression of GM-CSF mRNA. [Summary] VK2 acted on BM-MSCs to support their ability to enhance expansion and myeloid differentiation of BM-CD34+ cells probably via altered GM-CSF and CXCL12/SDF-1 expression in MSCs. These findings may help to identify the mechanisms of therapeutic effects of VK2 in patients with MDS (Figure). Disclosures No relevant conflicts of interest to declare.
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Su, Kuei-Ying, Takuya Nojima, Masayuki Kuraoka, M. Anthony Moody, and Garnett Kelsoe. "An in vitro culture system for studying the human B cell repertoire (P1459)." Journal of Immunology 190, no. 1_Supplement (May 1, 2013): 174.12. http://dx.doi.org/10.4049/jimmunol.190.supp.174.12.
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Abstract Defining the human B-cell repertoire is important for understanding human B-cell development and autoimmunity. Currently, re-expression of immunoglobulins from single B cells is the standard method to study human B-cell repertoire; however, this method is labor-intensive and costly as it requires large numbers of recombinant antibodies to be generated by single-cell PCR and expression of recombinant heavy and light chain pairs. We have developed an alternative method to characterize the B-cell repertoire by culture that supports the proliferation and differentiation of human pre-B, immature/transitional, and mature B cells recovered from bone marrow, fetal liver, or peripheral blood. Briefly, single B cells are cultured in IL-2, IL-4, IL-21, and BAFF on CD154+ stromal cells. This culture supports extensive cell proliferation and differentiation into IgG-secreting plasmacytes. The monoclonal IgG antibodies in each cell culture can be readily screened by ELISA, Luminex, or immunofluorescence against endogenous and exogenous antigens. We find that autoantibodies from pre-B cell cultures have higher avidities for self-antigens than do those from immature/transitional and mature B cell cultures, suggesting the loss of high avidity cells at the first tolerance checkpoint. V(D)J rearrangements are easily recovered from selected clones for sequencing and expression. This culture method offers a pathway to determine B-cell repertoires with high throughput and reduced cost.
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Su, Kuei-Ying, Masayuki Kuraoka, Akiko Watanabe, and Garnett Kelsoe. "Efficient culture of human naïve and memory B cells (LYM5P.711)." Journal of Immunology 194, no. 1_Supplement (May 1, 2015): 134.16. http://dx.doi.org/10.4049/jimmunol.194.supp.134.16.
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Abstract The ability to culture and expand B cell numbers in vitro has become a useful tool for studying human immunity. A limitation of current methods in human B cell cultures is the capacity to support continued proliferation by mature B cells. We have adapted the culture methods established by Luo et al. (Blood, 2009) to efficiently (60% cloning efficiency) activate and expand both naïve and memory human B cells. Briefly, naïve or memory B cells recovered from blood are cultured with cytokines on CD154+ feeder cells; this culture supports extensive B cell proliferation, with approximately 103 fold increases in a week; by splitting cultures onto new feeder layers, culture-derived (CD) B cells continuously proliferate, achieving 106 fold increases by day 16. The capacity for continued proliferation is stable for at least another week. In culture, a significant fraction of naïve B cells upregulate AID expression, undergo isotype switching without V(D)J hypermutation, and terminally differentiate into plasmacytes. CD B cells are readily cryopreserved and retain their ability to proliferate and differentiate. Significantly, proliferating CD B cells express high levels of MHCII, CD80, and CD86. We have examined the APC function of CD B cells and find that they present both allo- and microbial antigens to autologous T cells with comparable efficiency to PBMC. This culture method provides a platform for studying the BCR and TCR repertoires within a single individual.
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Jackson, Kevin S., Kari Inoue, David A. Davis, Tyvette S. Hilliard, and Joanna E. Burdette. "Three-Dimensional Ovarian Organ Culture as a Tool to Study Normal Ovarian Surface Epithelial Wound Repair." Endocrinology 150, no. 8 (May 7, 2009): 3921–26. http://dx.doi.org/10.1210/en.2008-1674.
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Ovarian cancers are primarily derived from a single layer of epithelial cells surrounding the ovary, the ovarian surface epithelium (OSE). Ovarian surface proliferation is associated with ovulation and has been suggested to play a role in ovarian surface transformation and cancer progression. Aspects of ovarian surface repair after ovulation include proliferation, migration, and surface regeneration. To study ovarian surface repair, an organ culture system was developed that supports the proliferation, encapsulation, and repair of an artificially wounded surface. Wounded mouse ovaries embedded into an alginate hydrogel matrix have normal OSE cells as demonstrated by expression of cytokeratin 8, vimentin, N-cadherin, and a lack of E-cadherin. Normal OSE cells began proliferating and migrating around wounded surfaces after 1 d of culture. Organ cultures were propagated in medium supplemented with BSA and fetal bovine serum to determine optimal growth conditions. BSA cultured organs had OSE that proliferated significantly more than controls until d 4, whereas fetal bovine serum cultured organs had significantly more surface area encapsulated by OSE. Overall, a three-dimensional ovarian organ culture supports the growth of normal OSE in response to artificial wounding and provides a novel system for investigating wound repair as it relates to the possible role of ovulation and ovarian cancer.
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Sahibzada, Khatera, Leslie B. Hammer, Margaret B. Neal, and Daniel C. Kuang. "The Moderating Effects of Work-Family Role Combinations and Work-Family Organizational Culture on the Relationship Between Family-Friendly Workplace Supports and Job Satisfaction." Journal of Family Issues 26, no. 6 (September 2005): 820–39. http://dx.doi.org/10.1177/0192513x05277546.
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This study determined whether work-family role combinations (i.e., work and elder care, work and child care, work and elder care and child care) and work-family culture significantly moderate the relationship between availability of workplace supports and job satisfaction. The data were obtained from the Families and Work Institute’s 1997 archival data set, the National Study of the Changing Workforce (NCSW). As predicted, the relationship between availability of workplace supports and job satisfaction varied depending on the type of work-family role combinations and levels of work-family culture. Specifically, the relationship was significant for the elder care work-family role combination, in that higher levels of workplace supports in unsupportive work-family cultures were associated with the greatest levels of job satisfaction. In addition, it was found that a supportive work-family culture and an increase in workplace supports were related to a slight decrease in job satisfaction for the elder care work-family role combination.
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Tisseret*, Brent, and Steven Vaughn. "Influence of Physical Parameters on the Growth, Morphogenesis and Volatile Monoterpene Production in Mentha spicata L. Cultures In Vitro." HortScience 39, no. 4 (July 2004): 778F—779. http://dx.doi.org/10.21273/hortsci.39.4.778f.
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The influence of altering the physical environment on the growth (fresh weight), morphogenesis (leaf, root, and shoot numbers) and secondary metabolism (i.e., volatile monoterpene, and carvone) of Mentha spicata L. (spearmint) shoots cultured on MS medium was studied. The type of physical support (e.g., agar, liquid, platforms, or glass supports) using Magenta vessels altered growth and morphogenesis. Mint shoots grown on liquid produced 4-x fold more fresh weight than on agar. Carvone levels were unaffected physical supports. Increasing the frequency of media replacement significantly increased growth without altering carvone. Vessel size influence was tested by culturing shoots on culture tubes, Magenta vessels and ½-gal. jars. Positive correlations occurred between vessel capacity and culture growth, morphogenesis and carvone levels. A comparative study testing several spearmint cultivars on either culture tubes or an automated plant culture system (APCS, a sterile hydroponics system) was conducted. The APCS produced more biomass (e.g., ≈15-x fold increase in fresh weight), morphogenesis and carvone than employing culture tubes. Carvone was only produced from shoots and was absent in either roots or callus. Carvone levels decreased proportionally in shoots as the distance from the shoot terminus increased. Altering the number of media culture immersions (4, 8, 12, or 16 immersions/day) with the APCS was tested. Twelve immersions of media/day was optimum. Higher culture growth rates resulted in lower carvone levels/culture; however, overall carvone levels/vessel increased due to greater biomass production.
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Hornick, J. E., F. E. Duncan, L. D. Shea, and T. K. Woodruff. "Multiple follicle culture supports primary follicle growth through paracrine-acting signals." REPRODUCTION 145, no. 1 (January 2013): 19–32. http://dx.doi.org/10.1530/rep-12-0233.
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In vitro follicle growth in alginate hydrogels is a unique and versatile method for studying ovarian and follicle biology that may also have implications for fertility preservation. Current culture systems support the development of isolated mouse follicles from the secondary stage onward. However, it has been a challenge to grow smaller follicles in vitro due to the dissociation of the oocyte from companion somatic cells. Recent work has demonstrated that coculturing primary follicles with mouse embryonic fibroblasts or ovarian stromal cells supports follicle survival and growth. In this study, we demonstrate that follicles themselves can exert a beneficial coculture effect. When primary follicles were cultured in groups of five or ten (multiple follicle culture), there was increased growth and survival. The multiple follicle culture approach maintained follicle integrity and resulted in the formation of antral stage follicles containing meiotically competent gametes. The growth and survival of primary follicles were highly number dependent, with the most significant enhancement observed when the largest number of follicles was grown together. Our data suggest that the follicle unit is necessary to produce the secreted factors responsible for the supportive effects of multiple follicle culture, as neither denuded oocytes, oocyte-secreted factors, nor granulosa cells alone were sufficient to support early follicle growth in vitro. Therefore, there may be signaling from both the oocyte and the follicle that enhances growth but requires both components in a feedback mechanism. This work is consistent with current in vivo models for follicle growth and thus advances the movement to recapitulate the ovarian environment in vitro.
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Subramaniam, Agatheeswaran, Mehrnaz Safaee Talkhoncheh, Kristijonas Zemaitis, Shubhranshu Debnath, Jun Chen, Mayur Jain, Roman Galeev, and Jonas Larsson. "Inhibition of Lysine-Specific Demethylase 1A (LSD1) Supports Human HSCs in Culture By Preserving Their Immature State." Blood 132, Supplement 1 (November 29, 2018): 194. http://dx.doi.org/10.1182/blood-2018-194.
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Abstract The molecular mechanisms that govern hematopoietic stem cell (HSC) fate decisions remain incompletely defined. It has been a long-standing goal in the field to gain a better understanding of the genes and pathways that regulate the self-renewal ability of HSCs in order to develop optimal culture conditions in which HSCs can be expanded for clinical benefit. Lysine-specific histone demethylase 1A (LSD1), also known as lysine (K)-specific demethylase 1A (KDM1A), regulates gene expression by specifically eliminating di- and mono-methyl groups on H3 lysine K4 and K9 residues. Studies in mice have shown that, conditional knockdown of LSD1 results in an expansion of bone marrow hematopoietic stem and progenitor cells (HSPCs). However, a complete knockout of LSD1 results in pancytopenia and a dramatic reduction of HSPCs. In this study, we asked whether inhibition of LSD1 would improve the maintenance or expansion of cultured human HSCs derived from umbilical cord blood (UCB). To evaluate the effect of LSD1 inhibition we treated UCB CD34+ cells with three different LSD1 inhibitors (2-PCPA, GSK-LSD1 and RN1) at their respective IC50 values (20µM, 16nM and 70nM) and expanded the cultures for 6 days in serum free medium supplemented with stem cell factor (SCF), thrombopoietin (TPO) and FMS-like tyrosine kinase 3 ligand (FLT3L). Since we (Subramaniam et. al. Haematologica 2018) and others recently have shown that EPCR is a reliable cell surface marker to track UCB derived HSCs during in vitro culture, we quantified the numbers of CD34+EPCR+ cells using flow cytometry and compared to DMSO treated control cultures. Remarkably, treatment with either 2-PCPA or GSK-LSD1 resulted in a more than 10-fold increase of CD34+EPCR+ cells, compared to controls. Further, from dose response experiments we found that 2-PCPA at 1.25 µM expanded the total CD34+ cell population more efficiently than GSK-LSD1, and we therefore used 2-PCPA at this concentration for the subsequent experiments. Using carboxyfluorescein succinimidyl ester (CFSE) labeling to monitor cell division, we found that 2-PCPA did not significantly alter the cell division rate of the cultured CD34+ cells compared to DMSO controls, suggesting that the expansion of CD34+EPCR+ cells is not due to increased proliferation, and that LSD1 inhibition rather may prevent differentiation of the immature HSPCs. To further explore this, we mapped the early transcriptional changes triggered by 2-PCPA in HSCs using gene expression profiling of CD34+CD38-CD45RA-CD90+ cells following 24 hours of culture with or without 2-PCPA treatment. We found that gene sets corresponding to UCB and fetal liver HSCs were significantly enriched upon 2-PCPA treatment compared to DMSO control (Normalized Enrichment Score (NES)=1.49, q=0.05). This suggest that 2-PCPA indeed restricts differentiation and preserves the HSC state upon ex vivo culture. Strikingly, the gene signature induced by LSD1 inhibition was highly similar to that induced by the known HSC expanding compound UM171 (NES=1.43, q=0.11). UM171 is a molecule with unknown target and has also been shown to dramatically expand the EPCR+ population in culture. Finally, the frequency of functional HSCs in DMSO and 2-PCPA treated cultures were measured using limiting dilution analysis (LDA). LDA was performed by transplanting 4 doses (day 0 equivalents of 20000, 1000, 300 and 100 CD34+ cells) of DMSO and 2-PCPA treated cultures into sub lethally irradiated (300cGy) NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice. Human CD45+ cell engraftment in the bone marrow was analyzed 18 weeks' post transplantation. Cultures treated with 2-PCPA showed a 5-fold higher content of long-term repopulating cells per day 0 CD34+ cell equivalent compared to the DMSO control (1 in 615 vs 1 in 3041, p=0.03). Thus, the 2-PCPA treated cultures had significantly enhanced HSCs numbers. To determine the absolute expansion rate, we are currently performing LDA using uncultured cells as well. Altogether our data suggest that LSD1 inhibition supports both phenotypic and functional HSCs in culture by preserving the immature state. Currently we are exploring the possibilities of using LSD1 inhibitors in combination with other known modifiers of HSC expansion. Disclosures No relevant conflicts of interest to declare.
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Prem, Caroline, and Bernd Pelster. "Swimbladder gas gland cells cultured on permeable supports regain their characteristic polarity." Journal of Experimental Biology 204, no. 23 (December 1, 2001): 4023–29. http://dx.doi.org/10.1242/jeb.204.23.4023.
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SUMMARY A cell culture system has been developed in which swimbladder gas gland cells from the European eel (Anguilla anguilla) were cultured on a permeable support. Cells seeded on Anodisc 13 (Whatman) or Costar Transwell 13 mm membranes form a confluent cell layer within the first 2 or 3 days of culture but, on the basis of measurements of transepithelial resistance, it is a ‘leaky’ cell layer. In a superfusion system, the apical and basal sides of the cells were superfused asymmetrically, with saline on the apical side and a glucose-containing cell culture medium on the basal side. Under these conditions, the cells continuously produced lactic acid, and approximately 60–70 % of this lactate was released at the basal side. To mimic the in vivo situation, the saline solution supplied to the apical side was replaced by humidified air in an additional series of experiments. Cells cultured in an air/liquid system produced even more lactate, and this lactate was only released to the basal side; there was no leakage of fluid to the apical side. After 4 or 5 days in the superfusion system, the cells were fixed for histological examination. The cells were columnar, similar to gas gland cells in vivo, and showed a clear polarity, with some small microvilli at the apical membrane and extensive membrane foldings at lateral and basal membranes. Immunohistochemical localization of Na+/K+-ATPase revealed that this ATPase was present mainly in the lateral membranes; it was never found in the apical membranes. Cells cultured in the air/liquid system showed a similar structure and polarity.
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Johnson, Adeerya. "Hella Bars: The Cultural Inclusion of Black Women’s Rap in Insecure." Open Cultural Studies 6, no. 1 (January 1, 2022): 76–87. http://dx.doi.org/10.1515/culture-2022-0144.
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Abstract The musical supervision of HBO’s insecure sonically maps various representations of Black women’s connections to hip-hop music as a site of autonomy, agency, and authenticity. Importantly, the variety of Black female rappers who are featured in seasons 1–3 of insecure connects nuanced and contemporary representations of Black millennial women’s understanding of Black womanhood, sex, friendship, love, and relationships. I argue that the influence of Issa Rae’s perception and connections to hip-hop and the placement of songs in insecure supports a soundtrack that takes on a hip-hop feminist approach to Black popular culture. I explore contemporary female hip-hop artist as an emerging group of rappers who support nuanced narratives and identities of Black millennial women. Furthermore, this article highlights the connectedness of Black popular culture and hip-hop feminism as an important site of representation for Black women who use hip-hop as a signifier to culture, self-expression, and identity. I recognize the importance of insecure’s soundtrack and usage of Black women in hip-hop to underline the ways hip-hop sits at the intersections of race, sexuality, and gender for Black women’s everyday lives.
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Zollers, Nancy J., Arun K. Ramanathan, and Moonset Yu. "The relationship between school culture and inclusion: How an inclusive culture supports inclusive education." International Journal of Qualitative Studies in Education 12, no. 2 (April 1999): 157–74. http://dx.doi.org/10.1080/095183999236231.
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Anggoro, Agung, and Ary Dwi Anjarini. "Building an Organizational Culture that Supports Diversity and Inclusion." Management Studies and Business Journal (PRODUCTIVITY) 1, no. 1 (January 31, 2024): 190–97. http://dx.doi.org/10.62207/12cjyv77.
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Building an organizational culture that supports diversity and inclusion is a significant challenge for organizations in this era of globalization. Through this research, we analyze the factors that influence the effectiveness of diversity and inclusion initiatives in organizations. The research results show that leadership commitment, inclusive policies, employee cultural awareness, employee involvement, and innovation play an important role in creating an inclusive and supportive work environment. Measuring the effectiveness of diversity and inclusion initiatives was also identified as an important step to ensure successful implementation. The practical and theoretical implications of these findings are discussed in the context of building an inclusive and supportive organizational culture.
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Endo, M., R. Kawahara-Miki, F. Cao, K. Kimura, T. Kuwayama, Y. Monji, and H. Iwata. "Estradiol supports in vitro development of bovine early antral follicles." REPRODUCTION 145, no. 1 (January 2013): 85–96. http://dx.doi.org/10.1530/rep-12-0319.
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Antrum formation and estradiol (E2) secretion are specific features of oocyte and granulosa cell complexes (OGCs). This study investigates the effect of E2 on the in vitro development of bovine OGCs derived from early antral follicles as well as on the expression of genes in granulosa cells (GCs). The supplementation of culture medium with either E2 or androstenedione (A4) improved the in vitro development of OGCs and the nuclear maturation of enclosed oocytes. When OGCs were cultured in medium containing A4, developmentally competent OGCs secreted more E2 than OGCs that were not competent. In addition, fulvestrant inhibited the effect of both E2 and A4 on OGCs development. Comprehensive gene expression analysis using next-generation sequence technology was conducted for the following three types of GCs: i) GCs of OGCs cultured for 4 days with E2 (1 μg/ml; E2(+)), ii) GCs of OGCs cultured for 4 days without E2 (E2(−)) or iii) OGCs that formed clear antrum after 8 days of in vitro culture in medium containing E2 (1 μg/ml; AF group). GCs of the E2(+) group had a similar gene expression profile to the profile reported previously for the in vivo development of large follicles. This genetic profile included factors implicated in the up-regulation of E2 biosynthesis and down-regulation of cytoskeleton and extracellular matrices. In addition, a novel gene expression profile was found in the AF group. In conclusion, E2 impacts the gene expression profile of GCs to support the in vitro development of OGCs.
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Habanjar, Ola, Mona Diab-Assaf, Florence Caldefie-Chezet, and Laetitia Delort. "3D Cell Culture Systems: Tumor Application, Advantages, and Disadvantages." International Journal of Molecular Sciences 22, no. 22 (November 11, 2021): 12200. http://dx.doi.org/10.3390/ijms222212200.
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The traditional two-dimensional (2D) in vitro cell culture system (on a flat support) has long been used in cancer research. However, this system cannot be fully translated into clinical trials to ideally represent physiological conditions. This culture cannot mimic the natural tumor microenvironment due to the lack of cellular communication (cell-cell) and interaction (cell-cell and cell-matrix). To overcome these limitations, three-dimensional (3D) culture systems are increasingly developed in research and have become essential for tumor research, tissue engineering, and basic biology research. 3D culture has received much attention in the field of biomedicine due to its ability to mimic tissue structure and function. The 3D matrix presents a highly dynamic framework where its components are deposited, degraded, or modified to delineate functions and provide a platform where cells attach to perform their specific functions, including adhesion, proliferation, communication, and apoptosis. So far, various types of models belong to this culture: either the culture based on natural or synthetic adherent matrices used to design 3D scaffolds as biomaterials to form a 3D matrix or based on non-adherent and/or matrix-free matrices to form the spheroids. In this review, we first summarize a comparison between 2D and 3D cultures. Then, we focus on the different components of the natural extracellular matrix that can be used as supports in 3D culture. Then we detail different types of natural supports such as matrigel, hydrogels, hard supports, and different synthetic strategies of 3D matrices such as lyophilization, electrospiding, stereolithography, microfluid by citing the advantages and disadvantages of each of them. Finally, we summarize the different methods of generating normal and tumor spheroids, citing their respective advantages and disadvantages in order to obtain an ideal 3D model (matrix) that retains the following characteristics: better biocompatibility, good mechanical properties corresponding to the tumor tissue, degradability, controllable microstructure and chemical components like the tumor tissue, favorable nutrient exchange and easy separation of the cells from the matrix.
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Sutkowski, N., ML Kuo, PS Amenta, JP Dougherty, and Y. Ron. "A peripheral blood-derived monolayer supports long-term cultures of human CD4+ and CD8+ T lymphocytes." Blood 85, no. 11 (June 1, 1995): 3213–22. http://dx.doi.org/10.1182/blood.v85.11.3213.bloodjournal85113213.
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An in vitro culture system has been developed for the long-term maintenance of primary, human peripheral blood and umbilical cord blood T lymphocytes, which does not rely on the use of stimulatory cytokines, antigen, or mitogens. In these cultures, a monolayer of adherent cells, some spindle-shaped and some resembling macrophages, developed within a week. All adherent cells were positive for the extracellular matrix proteins laminin and fibronectin, the intermediate filament vimentin, and for the surface markers major histocompatibility complex class II, platelet-endothelial cell adhesion molecule l (CD31), and E-Selectin (ELAM-1; CD62E). They were negative for the leukocyte common antigen (CD45), the macrophage marker MO-2 (CD14), muscle-specific actin, and Factor VIII-related antigen. These monolayers supported the maintenance of nonadherent, resting, mature T cells for up to 3 months, and these cells retained their ability to respond to mitogens and allogeneic cells. Both CD4+ and CD8+ cells were supported. The proportion of CD4+ and CD8+ cells remained unchanged after 3 months in culture. We have also used T cells from 2-month-old cultures as target cells for retroviral vector-mediated gene transfer. Up to 30% of the long-term T cells expressed the transferred lacZ gene after infection with a retroviral vector. The infection efficiency was similar to that obtained for fresh peripheral blood T cells, indicating that the long-term-cultured cells might be suitable for certain gene therapy applications.
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Sedlacek, Roger L., Ryan W. Carlin, Ashvani K. Singh, and Bruce D. Schultz. "Neurotransmitter-stimulated ion transport by cultured porcine vas deferens epithelium." American Journal of Physiology-Renal Physiology 281, no. 3 (September 1, 2001): F557—F570. http://dx.doi.org/10.1152/ajprenal.2001.281.3.f557.
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A collagenase-based dissociation technique has been developed to routinely establish monolayer cultures of freshly isolated porcine vas deferens epithelium. Cells isolated from each tissue are transferred to 25-cm2 tissue culture flasks and grown in a standard cell culture medium. Flasks reach confluency in 3–4 days, and cells are subsequently seeded onto permeable supports. Cultured cells display a monolayer cobblestone appearance and are immunoreactive to anti-ZO-1 and anti-cytokeratin antibodies. Electron microscopy is employed to demonstrate the presence of junctional complexes and microvilli. When evaluated in modified Ussing chambers, cultured monolayers exhibit a basal lumen negative potential difference, high electrical resistance (>1,000 Ω · cm2), and respond to norepinephrine, vasopressin, ATP, adenosine, and histamine, with changes in short-circuit current indicative of anion secretion. Responses are significantly attenuated in Cl−- and/or HCO[Formula: see text]-free solutions. Attempts to further optimize culture conditions have shown that chronic exposure to insulin increases proliferation rates. Thus the culture method described will reliably produce viable neurotransmitter-responsive cell monolayers that will allow for the characterization of vas deferens epithelial function and associated control mechanisms.
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Hosseini, Seyed Yaghoub, Khodakaram Salimifard, and Shahrbanoo Yadollahi. "On the Effects of Organizational Culture on E-Learning Readiness." International Journal of Social Ecology and Sustainable Development 3, no. 3 (July 2012): 42–52. http://dx.doi.org/10.4018/jsesd.2012070104.
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An organization’s success in implementing e-learning depends on the supports provided by the organizational culture. This paper is aimed to evaluate the impacts of organizational culture on e-learning readiness. To test the research hypothesis, Beta coefficient test was used. Research results indicated a significant positive impact of Clan and Adhocracy cultures on e-learning readiness. It was found that Market culture has a negative impact on e-learning readiness. Research findings cannot justify a relationship between Hierarchy culture and e-learning readiness.
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Nair, Arya Lekshmi, Lena Mesch, Ingo Schulz, Holger Becker, Julia Raible, Heiko Kiessling, Simon Werner, et al. "Parallelizable Microfluidic Platform to Model and Assess In Vitro Cellular Barriers: Technology and Application to Study the Interaction of 3D Tumor Spheroids with Cellular Barriers." Biosensors 11, no. 9 (September 3, 2021): 314. http://dx.doi.org/10.3390/bios11090314.
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Endothelial and epithelial cellular barriers play a vital role in the selective transport of solutes and other molecules. The properties and function of these barriers are often affected in case of inflammation and disease. Modelling cellular barriers in vitro can greatly facilitate studies of inflammation, disease mechanisms and progression, and in addition, can be exploited for drug screening and discovery. Here, we report on a parallelizable microfluidic platform in a multiwell plate format with ten independent cell culture chambers to support the modelling of cellular barriers co-cultured with 3D tumor spheroids. The microfluidic platform was fabricated by microinjection molding. Electrodes integrated into the chip in combination with a FT-impedance measurement system enabled transepithelial/transendothelial electrical resistance (TEER) measurements to rapidly assess real-time barrier tightness. The fluidic layout supports the tubeless and parallelized operation of up to ten distinct cultures under continuous unidirectional flow/perfusion. The capabilities of the system were demonstrated with a co-culture of 3D tumor spheroids and cellular barriers showing the growth and interaction of HT29 spheroids with a cellular barrier of MDCK cells.
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Kagan, Carolyn M., and Mark H. Burton. "Culture, identity and alternatives to the consumer culture." Educar em Revista, no. 53 (September 2014): 75–89. http://dx.doi.org/10.1590/0104-4060.36583.
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This article explores questions of identity and culture in relation to the present systemic crises that confront human life on the planet, problematizing the pursuit of economic growth and consumerist culture. It uses the concept of Ideology-Action-Structure complexes to understand the saturating nature of social, political and economic domination, and then explores interventions in these complexes, which all have characteristics of informal education, to promote cultural growth, create new settings and establish a counter-hegemonic ideology and alliance. It is suggested that by joining up fragmented local interventions and movements, there is hope that society's way of life can be shifted to one where cultural enrichment supports a less resource-exploitative economic and cultural model.
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Woodgate, Roberta L., and David Shiyokha Busolo. "African Refugee Youth’s Experiences of Navigating Different Cultures in Canada: A “Push and Pull” Experience." International Journal of Environmental Research and Public Health 18, no. 4 (February 20, 2021): 2063. http://dx.doi.org/10.3390/ijerph18042063.
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Refugee youth face challenges in navigating different cultures in destination countries and require better support. However, we know little about the adaptation experiences of African refugee youth in Canada. Accordingly, this paper presents the adaptation experiences of African refugee youth and makes recommendations for ways to support youth. Twenty-eight youth took part in semi-structured interviews. Using a thematic analysis approach, qualitative data revealed four themes of: (1) ‘disruption in the family,’ where youth talked about being separated from their parent(s) and the effect on their adaptation; (2) ‘our cultures are different,’ where youth shared differences between African and mainstream Canadian culture; (3) ‘searching for identity: a cultural struggle,’ where youth narrated their struggles in finding identity; and (4) ‘learning the new culture,’ where youth narrated how they navigate African and Canadian culture. Overall, the youth presented with challenges in adapting to cultures in Canada and highlighted how these struggles were influenced by their migration journey. To promote better settlement and adaptation, youth could benefit from supports and activities that promote cultural awareness with attention to their migration experiences. Service providers could benefit from newcomer-friendly and culturally sensitive training on salient ways of how experiences of multiple cultures affect integration outcomes.
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Luhur, Arthur, Daniel Mariyappa, Kristin M. Klueg, Kasun Buddika, Jason M. Tennessen, and Andrew C. Zelhof. "Adapting Drosophila melanogaster Cell Lines to Serum-Free Culture Conditions." G3: Genes|Genomes|Genetics 10, no. 12 (October 7, 2020): 4541–51. http://dx.doi.org/10.1534/g3.120.401769.
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Successful Drosophila cell culture relies on media containing xenogenic components such as fetal bovine serum to support continuous cell proliferation. Here, we report a serum-free culture condition that supports the growth and proliferation of Drosophila S2R+ and Kc167 cell lines. Importantly, the gradual adaptation of S2R+ and Kc167 cells to a media lacking serum was supported by supplementing the media with adult Drosophila soluble extract, commonly known as fly extract. The utility of these adapted cells lines is largely unchanged. The adapted cells exhibited robust proliferative capacity and a transfection efficiency that was comparable to control cells cultured in serum-containing media. Transcriptomic data indicated that the S2R+ cells cultured with fly extract retain their hemocyte-specific transcriptome profile, and there were no global changes in the transcriptional output of cell signaling pathways. Our metabolome studies indicate that there were very limited metabolic changes. In fact, the cells were likely experiencing less oxidative stress when cultured in the serum-free media supplemented with fly extract. Overall, the Drosophila cell culture conditions reported here consequently provide researchers with an alternative and physiologically relevant resource to address cell biological research questions.
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Ando, Kiyoshi, Yoshihiko Nakamura, Takashi Yahata, Yukari Muguruma, Tadayuki Sato, Hideyuki Matsuzawa, Tomoko Uno, Shunichi Kato, and Tomomitsu Hotta. "Angiopoietin-1 Supports SRC Activity Acquisition of Human CD34-Bone Marrow Cells." Blood 108, no. 11 (November 16, 2006): 1672. http://dx.doi.org/10.1182/blood.v108.11.1672.1672.
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Abstract CD34 negative hematopoietic stem cells (CD34− HSCs) were identified in mice and humans. Human HSCs are evaluated as severe combined immunodeficient mouse (SCID)-repopulating cells (SRCs), originally identified by the ability to reconstitute hematopoiesis in nonobese diabetic (NOD)/SCID mouse. CD34− cord blood (CB) cells have been hard to engraft in NOD/SCID mice until recent report of successlul engraftment by intra-bone marrow transplantation (iBMT). However, CD34− bone marrow (BM) cells have not been analyzed precisely. We prepared lineage negative CD34 negative (Lin-CD34−) cells by negative selection using CD2,3,7,14,16,19,20,33,34,36,41,56,127, and GlyA antibody. Lin-CD34− BM cells did not engraft in NOD/SCID mice even by using iBMT (0/6). In the previous study, we reported that Lin-CD34− BM cells were able to differentiate into CD34+ cells accompanied by the emergence of colony forming activity after 7 days of stroma-dependent culture, while SRC activity was not detected. (BMT 28, 587–595, 2001) Here we cultured Lin-CD34− BM cells on stroma cells transfected with human angiopoietin-1 cDNA (AHESS-5), since we detected Tie-2 expression on Lin-CD34− BM cells. AHESS-5 supported induction of CD34 much better than HESS-5 cells or empty vector transfected control cells (EVHESS-5), and the effect was blocked by anti-Tie-2 antibody (Fig.1). Furtheremore, CD34+ cells produced from CD34− BM cells engrafted in NOD/SCID mice (11/12). As previously reported, CD34− CB cells differentiate CD34+ cells and acquire SRC activity by stroma-dependent culture without angiopoietin-1. These results highlighted the characteristic differences of CD34− HSCs of BM from CB and the unique role of BM niche for CD34− HSCs. Fig. 1 CD34 expression on Lin − CD34 − BM cells after 7 days of culture Fig. 1. CD34 expression on Lin−CD34− BM cells after 7 days of culture
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Schock, Natalie, and Lieny Jeon. "ECE Program Supports and Teacher-Perceived Support from Families: Are They Connected?" Social Sciences 10, no. 10 (September 28, 2021): 361. http://dx.doi.org/10.3390/socsci10100361.
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According to the Conservation of Resources theory of stress, early care and education (ECE) teachers who receive greater tangible and interpersonal supports from their workplaces will be more positive and effective in their roles. This may translate to them perceiving or eliciting greater support from families, which is a key component to family engagement, a growing area of study in the ECE landscape. This study explores whether four program-level supports (benefits, professional development supports, teacher social supports, program-level family involvement activities) are associated with teacher-perceived support from families. The hypothesis was that all four will be positively associated. This study uses survey data from 102 preschool teachers and 13 preschool program directors in urban areas of two US states. We use ordinary least squares regression with cluster-robust standard errors and a stepwise build-up modeling procedure to determine associations between independent and dependent variables. While teacher social supports had the expected positive association with teacher-perceived support from families, family involvement activities were negatively associated. Our findings suggest that programs looking to improve family engagement may consider interpersonal/cultural supports for teachers and the larger school community. All else equal, simply offering more family involvement activities may not improve engagement culture.
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Johnston, Peter, Cheryl Dozier, and Julie Smit. "How Language Supports Adaptive Teaching Through a Responsive Learning Culture." Theory Into Practice 55, no. 3 (April 29, 2016): 189–96. http://dx.doi.org/10.1080/00405841.2016.1173992.
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Sailon, Alexander M., Alexander C. Allori, Edward H. Davidson, Derek D. Reformat, Robert J. Allen, and Stephen M. Warren. "A Novel Flow-Perfusion Bioreactor Supports 3D Dynamic Cell Culture." Journal of Biomedicine and Biotechnology 2009 (2009): 1–7. http://dx.doi.org/10.1155/2009/873816.
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Background. Bone engineering requires thicker three-dimensional constructs than the maximum thickness supported by standard cell-culture techniques (2 mm). A flow-perfusion bioreactor was developed to provide chemotransportation to thick (6 mm) scaffolds.Methods. Polyurethane scaffolds, seeded with murine preosteoblasts, were loaded into a novel bioreactor. Control scaffolds remained in static culture. Samples were harvested at days 2, 4, 6, and 8 and analyzed for cellular distribution, viability, metabolic activity, and density at the periphery and core.Results. By day 8, static scaffolds had a periphery cell density of , while in the core it was . Flow-perfused scaffolds demonstrated peripheral cell density of and core density of at day 8.Conclusions. Flow perfusion provides chemotransportation to thick scaffolds. This system may permit high throughput study of 3D tissues in vitro and enable prefabrication of biological constructs large enough to solve clinical problems.
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Shrove, Gina S., Ronald H. Olsen, and Timothy M. Vogel. "Development of pure culture biofilms ofP. putida on solid supports." Biotechnology and Bioengineering 37, no. 6 (March 15, 1991): 512–18. http://dx.doi.org/10.1002/bit.260370604.
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Goovaerts, I. G. F., J. L. M. R. Leroy, E. Merckx, S. Andries, and P. E. J. Bols. "210 CO-CULTURE WITH AUTOLOGOUS CUMULUS CELLS SUPPORTS THE INDIVIDUAL DEVELOPMENT OF SINGLY IN VITRO-MATURED AND FERTILIZED BOVINE OOCYTES." Reproduction, Fertility and Development 23, no. 1 (2011): 204. http://dx.doi.org/10.1071/rdv23n1ab210.
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The ability to produce embryos singly in vitro (in vitro production, IVP) would be a useful tool for many purposes. Without the interfering effects of other developing or degenerating oocytes or embryos, such an individual IVP system is the tool of choice for studies on oocyte quality and oocyte–embryo metabolism. Unfortunately, individual IVP in most cases leads to unsatisfactorily low blastocyst rates. Earlier work showed that individual culture of zygotes on a cumulus cell (CC) monolayer resulted in comparable numbers of good-quality embryos, as obtained following regular group culture (Goovaerts et al. 2009 Theriogenology 71, 729–738). However, co-culture with somatic cells is often criticised because of the undefined culture conditions and for sanitary reasons. In the cited study, CC for monolayer production were obtained from a different batch of ovaries. Our specific aim was to use CC from the zygote itself (autologous CC). Grade I COC (n = 660) were collected from slaughterhouse ovaries and randomly assigned to 2 treatments (5 replicates): a completely individual ‘single-oocyte’ IVP protocol, or routine group IVP as a control. Individual maturation (TCM-199 + 20% serum) and fertilization were performed in 20-μL droplets under oil in 24-well plates. Subsequently, each zygote was stripped and cultured in 20 μL of medium (SOF + 5% serum, 90% N2, 5% CO2, 5% O2), to which the autologous stripped CC were added. Group maturation and fertilization were carried out per 100 COC in 500 μL, whereas group culture was performed per 25 zygotes in 50-μL droplets under oil. Cleavage, blastocyst, and hatching rates were determined 2, 8, and 10 days post-fertilization, respectively. Possible effects of the individual and group cultures were evaluated with binary logistic regression (SPSS 15.0, SPSS Inc., Chicago, IL). No interactions between replicate and treatment were found (P > 0.05). Although a blastocyst rate of 15.1% was obtained using single IVP, the general efficacy of the single-embryo production system was lower when compared with group culture (Table 1). In conclusion, although developmental competence was impaired using individual IVP, co-culture with autologous cumulus cells can be useful in specific experimental setups in which the influence of other oocytes or embryos or heterologous somatic cells is unacceptable. Table 1.Cleavage, blastocyst, and hatching rates after individual and group in vitro production (IVP)
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Payne, Julianne, Laurie Cluff, Jason Lang, Dyann Matson-Koffman, and Antonio Morgan-Lopez. "Elements of a Workplace Culture of Health, Perceived Organizational Support for Health, and Lifestyle Risk." American Journal of Health Promotion 32, no. 7 (March 12, 2018): 1555–67. http://dx.doi.org/10.1177/0890117118758235.
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Purpose: We investigated the impact of elements of a workplace culture of health (COH) on employees’ perceptions of employer support for health and lifestyle risk. Design: We used 2013 and 2015 survey data from the National Healthy Worksite Program, a Centers for Disease Control and Prevention (CDC)-led initiative to help workplaces implement health-promoting interventions. Setting: Forty-one employers completed the CDC Worksite Health Scorecard to document organizational changes. Participants: Eight hundred twenty-five employees provided data to evaluate changes in their health and attitudes. Measures: We defined elements of a COH as environmental, policy, and programmatic supports; leadership and coworker support; employee engagement (motivational interventions); and strategic communication. Outcomes included scores of employees’ perceptions of employer support for health and lifestyle risk derived from self-reported physical activity, nutrition, and tobacco use. Analysis: We estimated effects using multilevel regression models. Results: At the employee level and across time, regression coefficients show positive associations between leadership support, coworker support, employee engagement, and perceived support for health ( P < .05). Coefficients suggest a marginally significant negative association between lifestyle risk and the presence of environmental and policy supports ( P < .10) and significant associations with leadership support in 2015 only ( P < .05). Conclusion: Relational elements of COH (leadership and coworker support) tend to be associated with perceived support for health, while workplace elements (environmental and policy supports) are more associated with lifestyle risk. Employers need to confront relational and workplace elements together to build a COH.
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Bakhsh Magsi, Hussain, Tze Ong, Jo Ho, and Ahmad Sheikh Hassan. "Organizational Culture and Environmental Performance." Sustainability 10, no. 8 (August 1, 2018): 2690. http://dx.doi.org/10.3390/su10082690.
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Because it has become more and more urgent for organizations to implement environmental strategies with the support of organizational culture, this study considers it necessary to conduct an empirical study to examine the impact of organizational culture on environmental performance. Synthesizing the perspectives of organizational culture and environmental performance, we applied a theoretical model in the manufacturing industry of Pakistan linking an organizational culture that supports environmental practices for better environmental performance. Based on a survey of 314 manufacturing firms, using Smart-PLS, the current study found that adaptability, mission and consistency positively affect environmental performance. However, involvement does not have an effect on environmental performance. Additionally, organizational culture as a latent variable has a strong impact on environmental performance. The study is one of the first, to the author’s knowledge that links OC and EP in a developing economy, in this case Pakistan.
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Tamir, Emanuel, and Sherry Ganon-Shilon. "A “cracking” school culture: leading resource exploitation during implementation of a national reform." Journal of Educational Administration 59, no. 5 (April 7, 2021): 650–65. http://dx.doi.org/10.1108/jea-09-2020-0198.
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PurposeThe study explores characteristics of strong school cultures through principals' exploitation of additional resources within implementation of a national reform.Design/methodology/approachAn interpretive approach was utilized to analyze qualitative data from semi-structured interviews with 35 Israeli high school principals who implemented a national reform in state and religious-state schools from all school districts.FindingsThe article presents four types of cracking cultures led by the principals: (1) a school values-based culture, such as respect; (2) a caring culture based on trust and a positive atmosphere; (3) a maintenance achievement-oriented culture; and (4) a creative culture that supports the teachers and takes risks in using resources beyond their intended purpose.Originality/valueExploring principals' exploitation of resources within a cracking culture may promote school improvement and innovation during national reform implementation.
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Ramming, David W., Richard L. Emershad, and Carol Foster. "In Vitro Factors During Ovule Culture Affect Development and Conversion of Immature Peach and Nectarine Embryos." HortScience 38, no. 3 (June 2003): 424–28. http://dx.doi.org/10.21273/hortsci.38.3.424.
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Various in vitro conditions for culture of ovules prior to extraction and culture of immature embryos of peach [Prunus persica (L.) Batsch] and nectarine [Prunus persica (L.) Batsch var. nucipersica Schneid.] were investigated. Culture vessels consisting of test tubes, petri dishes, and polycarbonate jars were tested along with various types of support and nutrient media. Agar support was superior to liquid media with filter paper supports. Agar produced the largest embryos with 90% to 93% being converted into plants compared to liquid with only 1% to 12% embryo conversion. The best ovule orientation and support was with the micropyle down and pushed halfway into an agar-gelled medium. In experiments two and three, test tubes with vertical ovule orientation (micropyle end of ovule pushed into agar) produced larger embryos, the largest plants and the greatest percentage of embryos that converted into plants (60% and 91%). Petri dish treatments were less successful in embryo conversion than test tubes and polycarbonate jars. The addition of activated charcoal (AC) to an agar-gelled medium produced significantly larger embryos with a similar conversion rate. The addition of an agar-gelled medium to culture vessels reduces preparation time compared to filter paper supports, and placing each ovule within a test tube eliminates cross contamination, making immature embryo culture more successful.
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KOPPADA, UMASHANKAR, PRADEEP MATAM, and GIRIDHAR PARVATAM. "In vitro elicitation supports the enrichment of 2H4MB production in callus suspension cultures of D. hamiltonii Wight & Arn." Romanian Biotechnological Letters 27, no. 1/2022 (January 10, 2022): 3302–8. http://dx.doi.org/10.25083/rbl/27.1/3302-3308.
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Influence of elicitation on in vitro production of 2-Hydroxy-4-Methoxy Benzaldehyde (2H4MB), a structural isomer of vanillin from callus suspension cultures of Decalepis hamiltonii was investigated. In vitro culture conditions were optimized to induce callus, suspension culture and biomass followed by metabolite production. Suspension cultures were established using leaf generated friable callus. Maximum content of 2H4MB production 0.079 ± 0.01 mg 100g-1 DW and biomass 197.5 ± 1.5 gL-1 were observed by 4th week of culturing. Elicitation was induced to suspension cultures by using m-topolin (mT), sodium nitroprusside (SNP), and pectin individually at different concentrations. m-topolin (15 μM), pectin (15 μM) and SNP (10 μM) supported 0.31 mg100g-1 DW, 0.27 mg100g-1 DW and 0.21 mg100g-1 of 2H4MB production respectively by 4th week. This data infers that the elicitation improves 2H4MB content in callus suspension cultures of D. hamiltonii.
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Kim, Jin. "Study on the Direction of Public Support for Metaverse." Korea Association Of Cultural Economics 25, no. 2 (August 31, 2022): 3–26. http://dx.doi.org/10.36234/kace.2022.25.2.3.
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In a situation where metaverse is regarded as a new benchmark of platform economies and vast investment on metaverse is needed in and out of country, this paper proposes the direction of public support for metaverse-related technologies based on the support direction of national R&D budget projects and the understanding of metaverse technology and market environments.
We investigate the current situation of R&D supports in the national level and admit the necessity of increasing CT(culture technology) investment. Furthermore, we find the R&D support principle that the direction of public supports in R&D investment be more desirable in indirect and ex post tax supports than in direct and ex ante fiscal supports, even in metaverse technology and market cases.
It is needed, firstly to increase tax supports for metaverse-related technologies by adding those to immersive contents or culture contents technologies in new-growth engine and proprietary technologies, and secondly to focus on global standardization for mega investment in the infrastructure area of metaverse value chains, and thirdly to promote small investment on creative and immersive innovations in the service area of metaverse value chains through deregulation and public private partnership. Therefore, in the short-run viewpoint and for the long-run performance of metaverse tech and market environments, the evaluation framework and tax expenditure evaluation is timely for the addition of metaverse-related technologies into new-growth engine and proprietary technologies in tax supports.
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Hasterok, Sylwia, Anna Gustafsson, and Anette Gjörloff Wingren. "Applications of Tumor Cells in an In Vitro 3D Environment." Applied Sciences 13, no. 18 (September 15, 2023): 10349. http://dx.doi.org/10.3390/app131810349.
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Spherical, multicellular aggregates of tumor cells, or three-dimensional (3D) tumor models, can be grown from established cell lines or dissociated cells from tissues in a serum-free medium containing appropriate growth factors. Air–liquid interfaces (ALIs) represent a 3D approach that mimics and supports the differentiation of respiratory tract and skin 3D models in vitro. Many 3D tumor cell models are cultured in conjunction with supporting cell types, such as fibroblasts, endothelial cells, or immune cells. To further mimic the in vivo situation, several extracellular matrix models are utilized to support tumor cell growth. Scaffolds used for 3D tumor cell culture growth include both natural and synthetic hydrogels. Three-dimensional cell culture experiments in vitro provide more accurate data on cell-to-cell interactions, tumor characteristics, drug discovery, metabolic profiling, stem cell research, and diseases. Moreover, 3D models are important for obtaining reliable precision data on therapeutic candidates in human clinical trials before predicting drug cytotoxicity. This review focuses on the recent literature on three different tissue types of 3D tumor models, i.e., tumors from a colorectal site, prostate, and skin. We will discuss the establishment of 3D tumor cell cultures in vitro and the requirement for additional growth support.
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Moyer, Patricia S. "Patterns and Symmetry: Reflections of Culture." Teaching Children Mathematics 8, no. 3 (November 2001): 140–44. http://dx.doi.org/10.5951/tcm.8.3.0140.
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Many contributions of diverse cultures foster a rich understanding of mathematics. Knowing how one's culture has contributed to mathematics and how these contributions enhance our cultural environment supports the acquisition of mathematical power. However, discussing culture in mathematics classrooms for a one-week celebration of women in mathematics or a one-month recognition of the contributions of African Americans is not enough. Cultural learning that recognizes race, ethnicity, gender, and social class should be woven into the fabric of mathematics lessons throughout the year. Yet many teachers have limited backgrounds in promoting culturally relevant mathematics in meaningful ways.
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Faucheux, C., S. Nesbitt, M. Horton, and J. Price. "Cells in regenerating deer antler cartilage provide a microenvironment that supports osteoclast differentiation." Journal of Experimental Biology 204, no. 3 (February 1, 2001): 443–55. http://dx.doi.org/10.1242/jeb.204.3.443.
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Deer antlers are a rare example of mammalian epimorphic regeneration. Each year, the antlers re-grow by a modified endochondral ossification process that involves extensive remodelling of cartilage by osteoclasts. This study identified regenerating antler cartilage as a site of osteoclastogenesis in vivo. An in vitro model was then developed to study antler osteoclast differentiation. Cultured as a high-density micromass, cells from non-mineralised cartilage supported the differentiation of large numbers of osteoclast-like multinucleated cells (MNCs) in the absence of factors normally required for osteoclastogenesis. After 48 h of culture, tartrate-resistant acid phosphatase (TRAP)-positive mononuclear cells (osteoclast precursors) were visible, and by day 14 a large number of TRAP-positive MNCs had formed (783+/−200 per well, mean +/− s.e.m., N=4). Reverse transcriptase/polymerase chain reaction (RT-PCR) showed that receptor activator of NF κ B ligand (RANKL) and macrophage colony stimulating factor (M-CSF) mRNAs were expressed in micromass cultures. Antler MNCs have the phenotype of osteoclasts from mammalian bone; they expressed TRAP, vitronectin and calcitonin receptors and, when cultured on dentine, formed F-actin rings and large resorption pits. When cultured on glass, antler MNCs appeared to digest the matrix of the micromass and endocytose type I collagen. Matrix metalloproteinase-9 (MMP-9) may play a role in the resorption of this non-mineralised matrix since it is highly expressed in 100 % of MNCs. In contrast, cathepsin K, another enzyme expressed in osteoclasts from bone, is only highly expressed in resorbing MNCs cultured on dentine. This study identifies the deer antler as a valuable model that can be used to study the differentiation and function of osteoclasts in adult regenerating mineralised tissues.
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Donahue, RE, YC Yang, and SC Clark. "Human P40 T-cell growth factor (interleukin-9) supports erythroid colony formation." Blood 75, no. 12 (June 15, 1990): 2271–75. http://dx.doi.org/10.1182/blood.v75.12.2271.2271.
Full textAbstract:
Abstract Because human P40 T-cell growth factor, tentatively designated interleukin-9 (IL-9), was isolated through its ability to stimulate a human IL-3-dependent leukemic cell line (M-O7E), we tested the ability of IL-9 to support the growth and differentiation of normal hematopoietic progenitor cells from peripheral blood and bone marrow. Although the M-O7E cell line was derived from a patient with megakaryoblastic leukemia, IL-9 has not proved to be a growth or maturation factor for megakaryocytes, but instead has proved to be effective in supporting the development of erythroid bursts (BFU-E) in cultures supplemented with erythropoietin. Using highly purified progenitors from peripheral blood, IL-3 showed a BFU-E plating efficiency of 46% compared with 20% for IL-9. Because of the purity of these cell preparations and the low cell density in culture, IL-9 is likely to interact directly with erythroid progenitors. Analysis of mixing experiments and of the morphology of the BFU-E in culture indicated that IL-9 interacts preferentially with a relatively early population of IL-3-responsive BFU-E. In cultures of human bone marrow or cord blood, IL-9 selectively supported erythroid colony formation, while IL-3 and granulocyte/macrophage colony-stimulating factor additionally yielded granulocyte/macrophage colonies. Therefore, IL-9 represents a new T cell-derived cytokine with the potential for selectively stimulating erythroid development in the hematopoietic system.
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Donahue, RE, YC Yang, and SC Clark. "Human P40 T-cell growth factor (interleukin-9) supports erythroid colony formation." Blood 75, no. 12 (June 15, 1990): 2271–75. http://dx.doi.org/10.1182/blood.v75.12.2271.bloodjournal75122271.
Full textAbstract:
Because human P40 T-cell growth factor, tentatively designated interleukin-9 (IL-9), was isolated through its ability to stimulate a human IL-3-dependent leukemic cell line (M-O7E), we tested the ability of IL-9 to support the growth and differentiation of normal hematopoietic progenitor cells from peripheral blood and bone marrow. Although the M-O7E cell line was derived from a patient with megakaryoblastic leukemia, IL-9 has not proved to be a growth or maturation factor for megakaryocytes, but instead has proved to be effective in supporting the development of erythroid bursts (BFU-E) in cultures supplemented with erythropoietin. Using highly purified progenitors from peripheral blood, IL-3 showed a BFU-E plating efficiency of 46% compared with 20% for IL-9. Because of the purity of these cell preparations and the low cell density in culture, IL-9 is likely to interact directly with erythroid progenitors. Analysis of mixing experiments and of the morphology of the BFU-E in culture indicated that IL-9 interacts preferentially with a relatively early population of IL-3-responsive BFU-E. In cultures of human bone marrow or cord blood, IL-9 selectively supported erythroid colony formation, while IL-3 and granulocyte/macrophage colony-stimulating factor additionally yielded granulocyte/macrophage colonies. Therefore, IL-9 represents a new T cell-derived cytokine with the potential for selectively stimulating erythroid development in the hematopoietic system.
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Sanchez, Cecilia G., Katie Hamel, Emma Rogers, Jordan Robinson, Haley Lassiter, Trivia Frazier, and Christopher Williams. "Abstract 4227: Human-derived hydrogels to support phenotypic changes in ER+ breast cancer cell line." Cancer Research 84, no. 6_Supplement (March 22, 2024): 4227. http://dx.doi.org/10.1158/1538-7445.am2024-4227.
Full textAbstract:
Abstract Adipose tissue plays a critical role in breast cancer incidence, progression, and response to therapy. The development of human derived 3D culture systems can accelerate the understanding of the role of environmental factors in the progression of Breast cancer as well as a model for preclinical studies during drug development. MCF-7 spheroids were cultured in Obatala Sciences’ Obavate and human-derived hydrogels ObaGel® and ObaGel®-ECM and control media. Organoid structure and phenotypic changes were analyzed via live cell imaging on the Incucyte S3. Migration studies were performed and gene expression analysis of EMT markers. Finally, adipocyte-breast cancer cell crosstalk was evaluated using pooled adipocytes co-cultured with MCF-7 spheroids. The results demonstrated that ObaGel® and ObaGel®-ECM 3D cultures support the growth and proliferation of MCF-7 tumorspheres during an extended culture period. In addition, MCF-7 cells exhibit the characteristic morphology of tumorsphere-forming cells when cultured in Obavate, with morphological changes in the ObaGel® and ObaGel®-ECM 3D. ObaGel® and ObaGel®-ECM differentially supports MCF-7 migratory behavior and phenotypic changes characteristic of EMT. Conclusions: Human derived hydrogels are developed to support 3D culture of breast cancer cells and to recapitulate the phenotypic changes associated with their metastatic potential. Furthermore, the use of a human derived 3D coculture system provides a platform for the understanding of adipose tissue-breast cancer cell interactions as it relates to breast cancer initiation, progression, and treatment response. Defining these interactions will support the development of a tool for future use in patient stratification and precision medicine in identifying underlying causes of breast cancer progression and response to treatments. Citation Format: Cecilia G. Sanchez, Katie Hamel, Emma Rogers, Jordan Robinson, Haley Lassiter, Trivia Frazier, Christopher Williams. Human-derived hydrogels to support phenotypic changes in ER+ breast cancer cell line [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4227.
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Daniele, Elena, Lorenzo Bosio, Noor Ahmed Hussain, Barbara Ferrari, Stefano Ferrari, Vanessa Barbaro, Brian McArdle, et al. "Denuded Descemet’s membrane supports human embryonic stem cell-derived retinal pigment epithelial cell culture." PLOS ONE 18, no. 2 (February 6, 2023): e0281404. http://dx.doi.org/10.1371/journal.pone.0281404.
Full textAbstract:
Recent clinical studies suggest that retinal pigment epithelial (RPE) cell replacement therapy may preserve vision in retinal degenerative diseases. Scaffold-based methods are being tested in ongoing clinical trials for delivering pluripotent-derived RPE cells to the back of the eye. The aim of this study was to investigate human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cells survival and behaviour on a decellularized Descemet’s Membrane (DM), which may be of clinical relevance in retinal transplantation. DMs were isolated from human donor corneas and treated with thermolysin. The DM surface topology and the efficiency of the denudation method were evaluated by atomic force microscope, scanning electron microscopy and histology. hESC-RPE cells were seeded onto the endothelial-side surface of decellularized DM in order to determine the potential of the membrane to support hESC-RPE cell culture, alongside maintaining their viability. Integrity of the hESC-RPE monolayer was assessed by measuring transepithelial resistance. RPE-specific gene expression and growth factors secretion were assessed to confirm maturation and functionality of the cells over the new substrate. Thermolysin treatment did not affect the integrity of the tissue, thus ensuring a reliable method to standardize the preparation of decellularized DM. 24 hours post-seeding, hESC-RPE cell attachment and initial proliferation rate over the denuded DM were higher than hESC-RPE cells cultured on tissue culture inserts. On the new matrix, hESC-RPE cells succeeded in forming an intact monolayer with mature tight junctions. The resulting cell culture showed characteristic RPE cell morphology and proper protein localization. Gene expression analysis and VEGF secretion demonstrate DM provides supportive scaffolding and inductive properties to enhance hESC-RPE cells maturation. Decellularized DM was shown to be capable of sustaining hESC-RPE cells culture, thus confirming to be potentially a suitable candidate for retinal cell therapy.
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Kismono, Gugup, and Raden Muhammad Pradana Ramadista. "The Effect of the Degree of Misfit Between Human Resources Management Practices and the Types of Organizational Culture on Organizational Performance." Gadjah Mada International Journal of Business 22, no. 3 (December 11, 2020): 301. http://dx.doi.org/10.22146/gamaijb.56583.
Full textAbstract:
“Fit model” argues that the level of misfit between human resources management (HRM) practices and the type of organizational culture negatively influences organizational performance. However, the lack of emprirical research to support that contention can be problematic. Utilizing the concept of fit, this study aims to examine empirically the effect of the degree of misfit between HRM practices and the types of organizational cultures on organizational performance. Data were collected from a sample comprising of 128 respondents representing 64 companies in Indonesia, from nine industrial sectors. The hypothetical model was developed based on four types of HRM practices (human relations, internal process, rational goals, and open systems) and four types of organizational cultures (clan, hierarchy, market, and adhocracy). Euclidean distance scores were calculated to describe the misfit between the HRM practices and the types of organizational culture variables. Subsequently, the effect of the misfit scores on organizational performance was determined. The results show that the degree of misfit between HRM practices and the type of organizational culture has a significant and negative effect on organizational performance. This empirical research supports the concept of fit, in which the type of organizational culture that is supported by suitable HRM practices will result in a more positive organizational performance. Then, it is deemed necessary for companies to adapt their HRM practices to their culture, in order to improve their performance.