Дисертації з теми "Primary cell culture of bivalve"
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Fleurbaix, Emmanuel. "Évaluation écotoxicologique des éléments terres-rares : approches cellulaires chez différentes espèces aquatiques." Electronic Thesis or Diss., Université de Lorraine, 2021. http://www.theses.fr/2021LORR0324.
Since 30 years ago, the growing use of Lanthanides in new technologies has contributed to important releases of these metals into aquatic ecosystems. In a global sustainable development policy aimed at preserving the quality of ecosystems, the impact of Lanthanides on aquatic organisms has naturally been questioned. However, studies on the aquatic ecotoxicology of Lanthanides are incomplete, and no consensus is established yet. In this context, we studied the cellular toxicity of Lanthanides individually and in mixtures. To determine these toxic effects, cell viability was measured on Danio rerio fibroblast-like cells (ZF4; ATCC®, CRL-2050™), Danio rerio hepatic cells (ZFL; ATCC®, CRL-2643™), Oncorhynchus mykiss epithelial cells (RTgill-W1; ATCC®, CRL-2523™), and primary culture of Corbicula fluminea digestive glands exposed to Lanthanides. Direct toxicity of Lanthanides has been observed on all cellular models. Concerning the toxicity of Lanthanides in mixtures, synergistic effects have been underlined on the three fish cell lines. In this research, we focused on the mechanisms of the detoxification of Lanthanides in the case of ZF4 cells from Danio rerio. The effects of Lanthanides were assessed in the presence of specific inhibitors of glutathione-S-transferases (ethacrynic acid) and MRP-like (MK571 and probenecid), by cell viability measurements. We decided to study these actors of the cellular detoxification due to their respective roles in phases II and III of the cellular detoxification of metals in fishes and bivalves. Regarding the results, MRP-like proteins are effectively involved in the detoxification of Lanthanides in ZF4 cells. Overall, our results highlighted the relevance of the toxic effects of Lanthanides at the cellular level for the risk assessment of these metals
Birmelin, Claudia. "Development of primary cell culture systems from marine invertebrates for use in toxicology." Thesis, University of Surrey, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.265684.
Bohm, Maren [Verfasser]. "The role of sialic acids in avian influenza virus infection of primary cell culture / Maren Bohm." Hannover : Bibliothek der Tierärztlichen Hochschule Hannover, 2010. http://d-nb.info/1004206291/34.
Tradewell, Miranda Lee. "The central role of calcium dysregulation in a primary cell culture model of amyotrophic lateral sclerosis." Thesis, McGill University, 2009. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=32355.
La sclérose latérale amyotrophique (SLA, alias la maladie de Lou Gehrig) est une maladie neuromusculaire à évolution rapide qui commence à l'âge adulte, et pour laquelle il existe présentement très peu de traitements. L'excitotoxicité du glutamate, la formation d'inclusions protéiques, la déficience du protéasome et le dysfonctionnement mitochondrial ont tous été associés à la SLA sporadique et héréditaire, mais on se questionne toujours sur ce qui rassemble ces éléments qui, ensemble, mènent au dysfonctionnement neuromusculaire et au décès. Dans les modèles de culture de SLA héréditaire causée par des mutations de l'enzyme Cu/Zn-superoxyde dismutase (SOD1), les traitements réduisant le Ca2+ intra-cellulaire prolongent la viabilité et empêchent la formation d'inclusions mutantes de la SOD1. Les motoneurones sont vulnérables aux surcharges de Ca2+ causées par une régulation du Ca2+ inadéquate et par un grand apport glutamatergique. Ainsi, la perturbation de l'homéostasie du calcium joue peut-être un rôle précoce important dans la SLA. Le but de cette thèse était d'enquêter sur la façon dont les niveaux de Ca2+ à l'intérieur des organites assurant la régulation du cytosol et du calcium (mitochondrie et réticulum endoplasmique) changent dans les motoneurones d'un modèle expérimental de la SLA, ainsi que sur la façon dont ces changements sont liés à d'autres marques de pathogénie de la SLA (détérioration du fonctionnement mitochondrial et du protéasome). Afin d'atteindre cet objectif, la G93A-SOD1 causant la SLA a été introduite dans les cultures de motoneurones provenant de neurones ganglions de la racine dorsale mu
Stab, II Bernd Robert. "The Effects of Cell Culture Oxygen Levels on the Replicative Senescence Processes of Primary Human Fibroblasts." Diss., Virginia Tech, 2009. http://hdl.handle.net/10919/28468.
Ph. D.
Hang, Ta-Chun. "Optimization of primary endothelial culture methods and assessment of cell signaling pathways in the context of inflammation." Thesis, Massachusetts Institute of Technology, 2012. http://hdl.handle.net/1721.1/71467.
Cataloged from PDF version of thesis.
Includes bibliographical references.
Tissue engineering is a potentially valuable tool for clinical treatment of diseases where host tissues or organs need to be replaced. Progression of engineering metabolically complex organs and tissues has been severely limited by the lack of established, functional vasculature. The thesis work described herein focused on methods of establishing and studying specific endothelial cell types in vitro for potential applications in establishing functional microvascular architecture. To achieve these objectives, a model system of primary liver sinusoidal endothelial cells (LSEC) was initially studied due to the high metabolic requirements of the liver, as well as the unique phenotype that they possess. We were able to demonstrate that free fatty acids were able to rescue LSEC in culture, promote proliferation, and maintain their differentiated phenotype. Our work with lipid supplementation in serum-free conditions provides flexibility in engineering liver tissue with a functional vasculature comprised with relevant endothelial types encountered in vivo. Following up our work with LSEC, we explored the human dermal microvascular endothelial cell (HDMVEC) system to understand the signaling mechanisms involved in sprouting angiogenesis. Engineered tissues that are implanted will require integration with host vasculature. We established a method to collect large signaling data sets from a physiologically relevant in vitro culture system of HDMVEC that permitted angiogenic sprouting. We were able to find statistically significant data regarding how angiostatic cues like Platelet Factor 4 can modulate angiogenesis signaling pathways. Our results from working with both types of endothelial cell systems provide insight into potential methods for establishing specialized microvasculature for engineered tissues, both in propagation of differentiated endothelial cells in vitro and promotion of tissue/organ survival following their implantation.
by Ta-Chun Hang.
Ph.D.
Kraft, Robert, Allon Kahn, José L. Medina-Franco, Mikayla L. Orlowski, Cayla Baynes, Fabian López-Vallejo, Kobus Barnard, Gerald M. Maggiora, and Linda L. Restifo. "A cell-based fascin bioassay identifies compounds with potential anti-metastasis or cognition-enhancing functions." The Company of Biologists, 2013. http://hdl.handle.net/10150/605272.
The actin-bundling protein fascin is a key mediator of tumor invasion and metastasis and its activity drives filopodia formation, cell-shape changes and cell migration. Small-molecule inhibitors of fascin block tumor metastasis in animal models. Conversely, fascin deficiency might underlie the pathogenesis of some developmental brain disorders. To identify fascin-pathway modulators we devised a cell-based assay for fascin function and used it in a bidirectional drug screen. The screen utilized cultured fascin-deficient mutant Drosophila neurons, whose neurite arbors manifest the 'filagree' phenotype. Taking a repurposing approach, we screened a library of 1040 known compounds, many of them FDA-approved drugs, for filagree modifiers. Based on scaffold distribution, molecular-fingerprint similarities, and chemical-space distribution, this library has high structural diversity, supporting its utility as a screening tool. We identified 34 fascin-pathway blockers (with potential anti-metastasis activity) and 48 fascin-pathway enhancers (with potential cognitive-enhancer activity). The structural diversity of the active compounds suggests multiple molecular targets. Comparisons of active and inactive compounds provided preliminary structure-activity relationship information. The screen also revealed diverse neurotoxic effects of other drugs, notably the 'beads-on-a-string' defect, which is induced solely by statins. Statin-induced neurotoxicity is enhanced by fascin deficiency. In summary, we provide evidence that primary neuron culture using a genetic model organism can be valuable for early-stage drug discovery and developmental neurotoxicity testing. Furthermore, we propose that, given an appropriate assay for target-pathway function, bidirectional screening for brain-development disorders and invasive cancers represents an efficient, multipurpose strategy for drug discovery.
Song, Miyeoun. "Organ and primary culture of medaka (Oryzias latipes) testis: Test systems for the analysis of cell proliferation and differentiation." Doctoral thesis, Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2003. http://nbn-resolving.de/urn:nbn:de:swb:14-1059039500031-89370.
Klingbeil, Maria Fátima Guarizo. ""Comparação de dois métodos de obtenção celular para cultura primária de queratinócitos bucais humanos"." Universidade de São Paulo, 2006. http://www.teses.usp.br/teses/disponiveis/85/85131/tde-17052007-144619/.
The therapeutic procedures frequently used in oral treatments for the pathological diseases are surgical, resulting in failures of the mucosal continuity.The possibility to obtain transplantable oral epithelia from an in vitro cell culture opens new utilization perspectives not only to where it comes from, but also as a reconstructive matherial for other parts of the human body, such as: urethra, epithelia corneo-limbal, cornea, ocular surface. Many researchers still use controversial methods for obtaining cells. It was therefore evaluated and compared the efficiency in both methods: enzimatic and direct explant to obtain oral keratinocytes from human oral mucosa. Fragments of intra oral epithelial tissues from healthy human subjects, undergoing dental surgeries, were donated to the research project. The keratinocytes were cultivated over a feeder-layer from a previously irradiated 3T3 Swiss albino fibroblasts. In this study it was compared the time needed in the cell obtaintion, the best cell amount between both methods, the life-span, the cell capacity to form an in vitro epithelia and its morphologic structure. The results in the accessment of both methods have shown the possibility to obtain keratinocytes from a small oral fragment, but at the same time we may verify the advantages and peculiar restrictions for each one of both analyzed methods.
Geyer, Simone [Verfasser], and S. [Akademischer Betreuer] Scholpp. "Establishment of a three-dimensional cell culture system to study tubular structures - A comparative study of neuronal differentiation in zebrafish and in 2D and 3D zebrafish primary cell culture / Simone Geyer. Betreuer: S. Scholpp." Karlsruhe : KIT-Bibliothek, 2015. http://d-nb.info/1112224580/34.
Mahat, Bimit. "The Effects of Hypoxia on Human Adipose Tissue Lipid Storage and Mobilization Functions: From Primary Cell Culture to Healthy Men." Thesis, Université d'Ottawa / University of Ottawa, 2017. http://hdl.handle.net/10393/36865.
Gaffuri, Anne-Lise. "Drosophila melanogaster, as a model system to study the cell biology of neuronal GPCRs." Thesis, Paris 5, 2012. http://www.theses.fr/2012PA05T063.
The type-1 cannabinoid receptor (CB1R), the neuronal receptor for the major psychoactive substance of marijuana, is one, of the most abundant G-protein coupled receptors in the mammalian central nervous system. CB1R is traditionally described as a presynaptic receptor that retrogradely regulates synaptic transmission. In addition to this now relatively wellcharacterized function, in the last two decades it has become widely recognized that endocannabinoid (eCB) actions in the brain are not limited to the regulation of neurotransmission at established adult synapses. Indeed, eCB and CB1R are now recognized to be involved in brain development at the synaptic, neuronal and network levels. However, precise mechanisms underlying these processes remain poorly described. Since cellular mechanisms that mediate CB1R-activition dependent neuronal remodeling and subneuronal targeting have been demonstrated to be cell-autonomous, we aimed to combine the power of Drosophila genetics with the experimental accessibility and single-cell resolution of lowdensity primary neuronal cultures, a tool currently lacking in Drosophila. Moreover, becauseDrosophila does not have a CB1R ortholog, CB1R cell biology may be observed independently from eCB machinery. Thus, we first developed and validated an in vitro culture protocol that yields mature and fully differentiated Drosophila neurons. Secondly, we showed that activation-dependent endocytosis of ectopically expressed CB1R is conserved in Drosophila neurons. Next, we investigated whether ectopic expression and activation of CB1R in Drosophila modulate neuronal development. As observed in mammals, we observed that activation of CB1R impairs dendritogenesis in a cell-autonomous manner. For further characterization of our model, we showed that, as with mammals, transient ectopic CB1R expression and activation in mushroom body neurons (the center of olfactory memory in Drosophila) modulate the formation of a consolidated form of aversive memory. In conclusion, the validation of this new animal model opens new perspectives to better characterize mechanisms underlying modulation of neuronal functions induced by CB1Ractivity
Guan, Haoji. "Double-stranded RNA induced gene silencing of neuropeptide genes in sand shrimp, Metapenaeus ensis and development of crustacean primary cell culture /." Click to view the E-thesis via HKUTO, 2006. http://sunzi.lib.hku.hk/hkuto/record/B36893626.
Tirnitz-Parker, Janina Elke Eleonore. "Primary culture and immortal cell lines as in vitro models to evaluate the role of TWEAK signalling in hepatic oval cells /." Connect to this title, 2007. http://theses.library.uwa.edu.au/adt-WU2008.0039.
Gonsalves, Kyle Joseph. "An exploration of RNA and miRNA expression and their role in cell cycle regulation of human primary trabecular meshwork cells." Thesis, University of Iowa, 2019. https://ir.uiowa.edu/etd/6744.
Daukste, Liene. "Mathematical Modelling of Cancer Cell Population Dynamics." Thesis, University of Canterbury. Department of Mathematics and Statistics, 2012. http://hdl.handle.net/10092/10057.
Chow, Sheung Ching. "The characterization of hyperosomotic stress-induced signaling cascades and the downstream effectors in primary gill cell culture of Japanese eels, Anguilla japonica." HKBU Institutional Repository, 2010. http://repository.hkbu.edu.hk/etd_ra/1182.
Abd, el Rahman Sahar El Sayed El Sayed Ali [Verfasser]. "Comparative analysis of current infectious bronchitis virus isolates in primary cell culture systems / Sahar El Sayed El Sayed Ali Abd El Rahman." Hannover : Bibliothek der Tierärztlichen Hochschule Hannover, 2010. http://d-nb.info/1009660683/34.
Stott, Lucy Claire. "Development and uses of a primary fish gill cell culture system to investigate the uptake, efflux and metabolism of pharmaceuticals in ecotoxicology." Thesis, King's College London (University of London), 2016. https://kclpure.kcl.ac.uk/portal/en/theses/development-and-uses-of-a-primary-fish-gill-cell-culture-system-to-investigate-the-uptake-efflux-and-metabolism-of-pharmaceuticals-in-ecotoxicology(4213e713-7048-441b-aa06-044e4baf768e).html.
Pitombo, Jonleno Coutinho Paiva [UNESP]. "Impacto de andrógenos na diferenciação e atividade de osteoclastos em cultura celular." Universidade Estadual Paulista (UNESP), 2016. http://hdl.handle.net/11449/138870.
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Os mecanismos de ação dos andrógenos sobre homeostase e regulação das células que participam do turnover ósseo em fêmeas ainda são pouco compreendidos. Este trabalho teve como principal objetivo avaliar a participação de andrógenos na diferenciação e atividade de osteoclastos in vitro. Células totais de medula óssea de fêmur e tíbia de camundongos fêmeas foram utilizadas como fonte de células precursoras de osteoclastos, sendo cultivadas utilizando-se α-MEM suplementado e em presença de RANK-L (30ng/mL) e M-CSF (50ng/mL). As células foram tratadas com testosterona (T) diidrotestosterona (DHT) e antagonistas de receptores de hormônios sexuais, como flutamida (FLU) e fulvestranto (FUL). O anastrozol (ANA) foi usado para inibição da enzima aromatase e o etanol (0,01%) foi utilizado como controle. Após cinco dias, as células foram fixadas, coradas com TRAP e contadas, considerando-se células TRAP-positivas com 3 ou mais núcleos. Para o ensaio de atividade, foram utilizadas placas revestidas com fosfato de cálcio inorgânico e a área de reabsorção foi calculada com o auxílio de software. O estágio de diferenciação osteoclástica foi avaliado por RT-qPCR e a modulação da expressão de receptores para hormônios sexuais foi avaliada por Western Blot. Os andrógenos (T e DHT) não exerceram efeitos sobre a diferenciação e atividade de osteoclastos (ANOVA; p>0,05). Por outro lado, os tratamentos com ANA, FLU e FUL, associados ou não a T, regularam positivamente a diferenciação e atividade de osteoclastos. A expressão gênica de RANK, Catepsina K, NFATc1 e β3 integrina não foi alterada pelos tratamentos propostos (ANOVA; p>0,05). Além disso, os tratamentos com T, DHT, FLU e FUL modularam a expressão proteica do receptor de andrógeno (AR) e dos receptores de estrógeno (ERα e ERβ) por Western Blot. Tomados em conjunto, nossos resultados indicam que os andrógenos exercem limitada participação na diferenciação e atividade de osteoclastos de camundongos fêmeas e que este processo é mediado, ao menos em parte, por ações indiretas da T e pela modulação de receptores de hormônios sexuais.
The action mechanisms of androgens on homeostasis and the regulation of cells that participate in bone turnover in females are still poorly understood. This study had as main objective to evaluate the participation of androgens in the differentiation and activity of in vitro osteoclasts. Total bone marrow cells from femur and tibia of female mice were used as a source of precursor cells of osteoclasts, they were cultivated using supplemented α-MEM and in the presence of RANK-L (30ng/mL) and M-CSF (50ng/mL). The cells were treated with testosterone (T), dihydrotestosterone (DHT) and antagonists of sexual hormone receptors such as flutamide (FLU) and fulvestrant (FUL). Anastrozole (ANA) was used for inhibiting the aromatase enzyme and ethanol (0.01%) was used as a control. After five days, the cells were fixed, colored with TRAP and counted, considering TRAP-positive cells those ones containing 3 or more nuclei. For the activity assay, were used plaques covered with inorganic calcium phosphate and the area of reabsorption was calculated with the assistance of a software. The osteoclast differentiation stage was evaluated by RT-qPCR and the modulation of the expression of receptors for sexual hormones was assessed by Western Blotting. The androgens (T and DHT) did not exert effects in differentiation and activity of osteoclasts (ANOVA, p> 0.05). On the other hand, the treatments with ANA, FLU and FUL, associated or not to T, positively regulated the differentiation and activity of osteoclasts. The genic expression of RANK, Cathepsin K, NFATc1 and β3 integrin was not altered by the proposed treatments (ANOVA, p> 0.05). Moreover, the treatments with T, DHT, FLU and FUL modulated the protein expression of the androgen receptor (AR) and of the estrogen receptors (ERα and ERβ) by Western Blotting. Taken together, our results indicate that the androgens exert limited participation in differentiation and activity of osteoclasts of female mice and that this process is mediated, at least in part, by indirect actions of T and by the modulation of sexual hormone receptors.
CNPq: 133815/2014-5
FAPESP: 2013/12014-6
Iji, Oluwafikemi Temitayo. "In vitro bioassays as tools for evaluating toxicity of acidic drainage from a coal mine in Mpumalanga, South Africa." Thesis, University of Pretoria, 2016. http://hdl.handle.net/2263/60126.
The study site was an impacted stream located downstream of a coal mine discharge point whose effluent flowed away from the mine. Water chemistry results suggested high AMD impact evidenced by acidity, elevated sulphates, increased conductivity and presence of heavy metals. Al, Fe, Zn, Mn and Si were the major metals of potential concern in the AMD impacted stream; sulphates and major ions like Ca, K, Na and Mg were present at levels above target water quality range (TWQR) for effluents in receiving stream. The AMD impacted stream caused increased generation of reactive oxygen species (ROS) detectable in vitro in selected cell lines (Vero, C3A and RTgill-W1 cell lines), an indication of oxidative stress. In-stream, active treatment with caustic soda was efficient at reducing metal burden, with subsequent reduction in ROS generation in fish gill cell lines. For in vitro cytotoxicity tests, passive and active treated AMD water was cytotoxic to cell lines (Vero and RTgill-W1), with the fish RTgill-W1 cells exhibiting greater sensitivity compared to the mammalian Vero cells. Mitochondria played a larger role in observed loss in cellular viability (increased vacuolization, mitochondrial membrane swelling and damage), which was detected using mitochondrial specific stains, and by transmission electron microscopy (TEM). Increased dose- dependent cytotoxicity was observed in the fish gill and mammalian cell lines. Cells exposed to water samples (AMD and reference sites) revealed significant differences (p < vi 0.05) between the AMD impacted watershed and a relatively pristine site (reference site) where exposure to the same cells maintained approximately 100% viability at all concentrations for up to 72h exposure. The observed differences in effect in this study demonstrate that the effluent from the coal mine negatively impacted surface water quality, resulting in toxicity to cell lines, therefore creating an environment that would not be conducive for the survival of biological aquatic communities and potentially of concern for downstream human end users.
The induction of cytochrome P450 (CYP) 1A and resultant increase in 7- ethoxyresorufin-o-deethylase activity in primary fish gill cultures exposed to polycyclic aromatic hydrocarbons B[a]P, a known AhR agonist contaminant associated with coal mining, showed that there was as increase in EROD activity which was not observed using the RTgill-W1 cell lines. Gill epithelial cells isolated from the gills of Tilapia fish (Oreochromis mossambicus) bear close similarities to fish gills in vivo and their capacity to respond to the presence of AhR indicates that they may serve as a simple, cost-effect screening tool for assessing PAHs and dioxin-like compounds in fresh water.
For genotoxicity evaluation, the Ames test performed without metabolic activation using bacterium Salmonella typhimurium TA98 and TA100 strains revealed no indication of genotoxic activity in any of the water samples. Genotoxicity assessment of all water samples using the comet assay however exposed DNA damage to Vero and RTgill-W1 cell lines. A significant reduction in DNA damage was observed following active treatment. The results suggest that neither treatment technologies employed were efficient at removing all potential genotoxicants so further improvements are required. The comet assay proved sensitive enough to detect genotoxicity in reference water samples despite no known untoward effluent inputs at the site, suggesting potential for this assay to be integrated into an environmental monitoring framework.
The results obtained support the use of in vitro bioassays for evaluating toxicity of industrial effluent through biological responses in test systems elicited following exposure, improving ability to detect AMD polluted water. This could be beneficial when assessing the degree and extent of impact of AMD in natural water sources, and the possible environmental impact resulting from hazardous elements present in effluent water. In conclusion, these results suggest that in vitro techniques involving cell lines and primary cultures from fish may serve vii as simple, rapid and cost-effective tools for assessing risk and potential toxic effects of contaminants in AMD waters.
Thesis (PhD)--University of Pretoria, 2016.
The National Research Foundation
Department of Paraclinical Sciences (University of Pretoria)
Schlumberger Stichting Fund, Netherlands
Paraclinical Sciences
PhD
Unrestricted
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.
Nakajima, Aya. "Radiation sensitivity assay with a panel of patient-derived spheroids of small cell carcinoma of the cervix." Kyoto University, 2015. http://hdl.handle.net/2433/199178.
Okolicsanyi, Rachel K. "Mesenchymal stem cells as mediators of the neuronal cell niche." Thesis, Queensland University of Technology, 2015. https://eprints.qut.edu.au/84485/1/Rachel_Okolicsanyi_Thesis.pdf.
Pinello, Katia Cristina. "Avaliação da quimiosensibilidade de mastocitomas caninos graus I, II e III ao ácido retinóico todo-trans." Universidade de São Paulo, 2006. http://www.teses.usp.br/teses/disponiveis/10/10133/tde-02032007-090228/.
Mast cell tumor (MCT) is one of the most frequent neoplasms that affect the skin and soft tissue of the dog, representing about 7% a 21% of all skin tumors and 11% a 27% of malignant skin tumors in this specie. They present a great variety of appearance and behavior, which becomes a challenge to the treatment. The retinoids are well recognized as promising antitumor agents. However, there have only been a few reports about the effect of retinoids in canine cancers. The aim of this study was to characterize the primary mast cell tumor culture and to investigate the chemosensitivity of this tumor to all trans retinoic acid (ATRA). The primary cell culture of MCT was performed as co-cultive with fibroblasts, showing a positive interaction between mast cells and fibroblasts, with a lifetime of 30 days. The chemosensitivity of MCT to ATRA showed no difference between grade II or III, thus either a MCT grade II or grade III has the same response with ATRA at the doses studied. It has been shown that the MCT is more sensible at the dose 10-4M (p < 0,002). There is also an effect on first 24h untill 48h, changing after 72h. According to these results, it is possible to state that the great chemosensitivity of MCT to ATRA is after 72h of exposition at 10-4M. We can conclude that ATRA may be a potential adjunctive chemotherapeutic agent for the treatment of canine mast cell tumor.
Khaghani, Seyed A. "Cell and tissue engineering of articular cartilage via regulation and alignment of primary chondrocyte using manipulated transforming growth factors and ECM proteins. Effect of transforming growth factor-beta (TGF-¿1, 2 and 3) on the biological regulation and wound repair of chondrocyte monolayers with and without presence of ECM proteins." Thesis, University of Bradford, 2010. http://hdl.handle.net/10454/5371.
Pitombo, Jonleno Coutinho Paiva. "Impacto de andrógenos na diferenciação e atividade de osteoclastos em cultura celular /." Araraquara, 2016. http://hdl.handle.net/11449/138870.
Banca: Ticiana Sidorenko de Oliveira Capone
Banca: Thallita Pereira Queiroz
Resumo: Os mecanismos de ação dos andrógenos sobre homeostase e regulação das células que participam do turnover ósseo em fêmeas ainda são pouco compreendidos. Este trabalho teve como principal objetivo avaliar a participação de andrógenos na diferenciação e atividade de osteoclastos in vitro. Células totais de medula óssea de fêmur e tíbia de camundongos fêmeas foram utilizadas como fonte de células precursoras de osteoclastos, sendo cultivadas utilizando-se α-MEM suplementado e em presença de RANK-L (30ng/mL) e M-CSF (50ng/mL). As células foram tratadas com testosterona (T) diidrotestosterona (DHT) e antagonistas de receptores de hormônios sexuais, como flutamida (FLU) e fulvestranto (FUL). O anastrozol (ANA) foi usado para inibição da enzima aromatase e o etanol (0,01%) foi utilizado como controle. Após cinco dias, as células foram fixadas, coradas com TRAP e contadas, considerando-se células TRAP-positivas com 3 ou mais núcleos. Para o ensaio de atividade, foram utilizadas placas revestidas com fosfato de cálcio inorgânico e a área de reabsorção foi calculada com o auxílio de software. O estágio de diferenciação osteoclástica foi avaliado por RT-qPCR e a modulação da expressão de receptores para hormônios sexuais foi avaliada por Western Blot. Os andrógenos (T e DHT) não exerceram efeitos sobre a diferenciação e atividade de osteoclastos (ANOVA; p>0,05). Por outro lado, os tratamentos com ANA, FLU e FUL, associados ou não a T, regularam positivamente a diferenciação e atividade d... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: The action mechanisms of androgens on homeostasis and the regulation of cells that participate in bone turnover in females are still poorly understood. This study had as main objective to evaluate the participation of androgens in the differentiation and activity of in vitro osteoclasts. Total bone marrow cells from femur and tibia of female mice were used as a source of precursor cells of osteoclasts, they were cultivated using supplemented α-MEM and in the presence of RANK-L (30ng/mL) and M-CSF (50ng/mL). The cells were treated with testosterone (T), dihydrotestosterone (DHT) and antagonists of sexual hormone receptors such as flutamide (FLU) and fulvestrant (FUL). Anastrozole (ANA) was used for inhibiting the aromatase enzyme and ethanol (0.01%) was used as a control. After five days, the cells were fixed, colored with TRAP and counted, considering TRAP-positive cells those ones containing 3 or more nuclei. For the activity assay, were used plaques covered with inorganic calcium phosphate and the area of reabsorption was calculated with the assistance of a software. The osteoclast differentiation stage was evaluated by RT-qPCR and the modulation of the expression of receptors for sexual hormones was assessed by Western Blotting. The androgens (T and DHT) did not exert effects in differentiation and activity of osteoclasts (ANOVA, p> 0.05). On the other hand, the treatments with ANA, FLU and FUL, associated or not to T, positively regulated the differentiation and activity ... (Complete abstract click electronic access below)
Mestre
Monteil, Christelle. "Modulation du phénotype exprimé par des cellules tubulaires proximales rénales en culture : stratégie pour le développement de modèles d'études in vitro en physiopathologie et pharmacotoxicologie rénales." Rouen, 1994. http://www.theses.fr/1994ROUES060.
Khaghani, Seyed Ali. "Cell and tissue engineering of articular cartilage via regulation and alignment of primary chondrocyte using manipulated transforming growth factors and ECM proteins : effect of transforming growth factor-beta (TGF-β1, 2 and 3) on the biological regulation and wound repair of chondrocyte monolayers with and without presence of ECM proteins". Thesis, University of Bradford, 2010. http://hdl.handle.net/10454/5371.
Udo, Mariana Sayuri Berto. "Caracterização da via de ativação de neurotoxicidade induzida pela Anidroegconina Metil Éster (AEME) in vitro." Universidade de São Paulo, 2017. http://www.teses.usp.br/teses/disponiveis/9/9141/tde-22012018-103721/.
Cocaine market is increasing all around the world. In Brazil it is estimated that almost 2 million people make usage of this substance which 370 thousand people use the crack form. Cocaine is a psychostimulant with large potential for abuse and the smokable form produces more health problems than the other routes of use, mainly in the cognitive field related to compromising attention, memory and decision take. The crack users are exposed to both volatized cocaine and their pyrolysis products, which the main product is the anhydroecgonine methyl ester (AEME). Considering that the cognitive disturbs could be related to neurons death, the memory functions are also related to the hippocampal functions, and little is known about the AEME neurotoxicity or even the combination of cocaine and AEME in cell fate, our study aims to characterize the time and pathways related to the hippocampal neurotoxicity induced by 2 mM of cocaine, 1 mM of AEME and the association (C + A) of both substances during 3 h, 6 h and 12 h of exposure. Our results showed that cocaine and AEME increased enzymatic activity (MTT test) in 3 h but it reversed during 12 h of exposure. Moreover, AEME increased cell permeability in 6 h keeping it until 12 h. Although theses early alterations, both substances activated caspase -8 after 12 h when early apoptosis was also observed by the FS externalization. Cocaine activated the autophagic process at 3 h increasing the LC3 II quantification, but decreased the number of cell with acid vesicle at 6 h and 12 h, suggesting neuronal death due to failure in the autophagic flux. AEME showed increased in cell number with acid vesicle only in 3 h which returned after 6 h suggesting that the autophagic process gave place to the apoptotic program starting from the extrinsic pathway. The association of cocaine and AEME was shown more neurotoxic than them alone, decreasing the number of integral cells after 3 h, activating caspase -8 and promoting FS externalization after 6 h without involving the autophagy. In addition, taking the C + A morphology in 6 h, where it was observed increasing of nucleus and soma size that became pyknotic at 12 h, we suggest that the neuronal death could occur by necroptosis because this composition activated caspase -8 and resulted in necrotic like morphology. Thus, we conclude that cocaine- and AEME-induced apoptosis neuronal death starts in 12 h of exposure by the extrinsic pathway and the association of both substances is more neurotoxic than they alone, starting earlier after 6 h and resulting in a necrotic-like morphology.
Franke, Jana, Vanessa Abs, Claudia Zizzadoro, and Getu Abraham. "Comparative study of the effects of fetal bovine serum versus horse serum on growth and differentiation of primary equine bronchial fibroblasts." Universitätsbibliothek Leipzig, 2014. http://nbn-resolving.de/urn:nbn:de:bsz:15-qucosa-144893.
Wölffling, Sarah. "Generation of a stem cell driven in vitro culture of polarized cells to study gastric tissue homeostasis and response to infections." Doctoral thesis, Humboldt-Universität zu Berlin, 2020. http://dx.doi.org/10.18452/21716.
In the human gastric mucosa, multiple interactions between different cell populations regulate digestion and surveillance of infections. Epithelial cells in the mucosa differentiate into specialized cell types to produce protective mucins, gastric acid, digestive enzymes or hormones. Infection with Helicobacter pylori dysregulates the tissue homeostasis increasing the chance to develop a gastric ulcer, adenocarcinoma or ultimately gastric cancer at the site of infection. In this thesis, the development of a novel in vitro culture model for human primary gastric epithelial cells, called the mucosoid culture, is shown. The mucosoid cultures are representative of epithelial barriers and recapitulate most of the functions of the human gastric mucosa in vivo, including mucus production, and allow long-term and stable cultivation of epithelial cells as well as infection studies with H.pylori. Corpus derived mucosoids were used to investigate the niche factors that promote the differentiation of foveolar cells, chief cells, and parietal cells. EGF was found to be a major regulator in differentiation together with BMP/Noggin. Stromal cells are part of the lamina propria of the gastric mucosa. Very little is known about the interaction with the epithelium under homeostatic conditions and during bacterial infections with Helicobacter pylori. The co-culture of human primary gastric stromal cells with epithelial cells using the mucosoid culture model demonstrated the active signaling between both cell types. Furthermore, mucosoid cultures were successfully infected with H. pylori. The results revealed that stromal cells actively respond to epithelial infection with cytokine and chemokine expression. Concurrently stromal cells increased the NFκB-driven inflammatory response in epithelial cells.
Grasso, Sonia. "Effect of growth factors, steroids, alfa-lipoic acid and Astroglial Conditioned Media on the expression of some biomarkers of astroglial cell proliferation and differentiation in primary culture." Doctoral thesis, Università di Catania, 2013. http://hdl.handle.net/10761/1323.
Brodlie, Malcolm James. "Development of a primary airway epithelial cell culture model and explanted tissue archive to study the role of neutrophilic inflammation and airway remodelling in cystic fibrosis lung disease." Thesis, University of Newcastle Upon Tyne, 2011. http://hdl.handle.net/10443/1228.
Cardoso, Matheus Völz. "Efeitos da fotobioestimulação por laser e LED nas células da granulação óssea." Universidade de São Paulo, 2017. http://www.teses.usp.br/teses/disponiveis/25/25146/tde-28092017-193857/.
Photobiomodulation by laser and LED is a new therapeutic non-invasive trend. Photophysical and photochemical effects occur in immunomodulation, acceleration of wound healing and angiogenesis and reduction of pain. These effects are desired in bone tissue but there are no defined parameters for light irradiation and no consensus for the best effect on osseous cells. The aim of this study was to evaluate photobiomodulation effects on cell viability and mineralization events of rat osseous granulation cells (rGO). Cells in 6th passage were plated in 96-well plates for viability tests (1x10³ cells), and 24-well plates for in vitro wound healing test (1x104 cells), mineralization and alkaline phosphatases (AF) activity (4x104 cells). Cells were cultured in DMEM (10% bovine fetal serum) and irradiation with lasers (AlGaAs-660nm e AlGaInP-810nm) and LED (637±15nm). Experimental groups were red laser (3 and 5 J/cm²), infrared laser (3 and 5 J/cm²), LED (3 and 5s), positive(C+) and negative controls (C-, 1% bovine fetal serum). For mineralization and AF assays, other groups with osteogenic medium and same light treatments were added. Cell viability was evaluated by MTT and crystal violet tests at 24, 48, 72 and 96h. In vitro wound healing test evaluated the percentage of wound closure area by cells migration at 12, 24, 36, 48h. Mineralization test was done by alizarin red at 14, 21 and 28 days. AF activity was measured at 7, 14 and 21 days. Statistical analysis was performed by ANOVA complemented by Tukeys test (p<0,05). Results showed that light therapies in general increased viability and wound healing closure, mostly red laser and LED5s (p<0,05). Best results in mineralization stimulation were observed for LED5s. In groups with osteogenic medium, a synergistic effect of photobiomodulation resulted in higher numbers of mineral nodules. Light groups stimulated higher mineral nodule formation than positive control (p>0.05) even in groups with regular medium, showing an osteogenic induction by light. Increased AF activity was observed at 7 days in light treatment groups (p<0,05). In conclusion, laser and LED photobiostimulation increased viability, cell migration and mineralization events in osteoblasts with best results for red laser and LED.
Hall-Ponselè, Andrew M. "Genetic engineering of the primary/secondary metabolic interface in tobacco BY-2 cells." Thesis, University of Oxford, 2014. http://ora.ox.ac.uk/objects/uuid:be5a3ee3-33c7-455c-b043-409987395f98.
Ha, Thi Binh Minh. "Contributions à l'étude des méthodes de production de masse des cellules endothéliales cornéennes humaines." Thesis, Saint-Etienne, 2014. http://www.theses.fr/2014STET001T/document.
Corneal endothelial engineering is becoming a more and more realistic solution to restore vision from corneal edema. This method focus to regenerate corneal endothelium by direct injection of corneal endothelial cells (ECs) into patient anterior chamber at the early stage of endothelial dystrophies, or by grafting a transparent biocompatible material covered by a monolayer of ECs. These two techniques require both in vitro isolation and amplification of ECs or endothelial-like cells. In this thesis, different strategies to obtain a high quantity of functional ECs for clinical application are explored: 1- Due to the limit proliferative capacity of EC, the first strategy consists to analyze mechanisms implicated EC cell cycle arrest and then to optimize protocol for native EC isolation or for cell proliferation activation ex vivo. This is summarized in three publications. The first publication describes the cell cycle regulation by comparing transcriptional expression of 112 genes in 6 biological models of EC with different proliferative profile: in vivo, postmortem, organ-culture, confluent primary culture, non confluent primary culture and immortalized cell line. , The key molecular actors identified using the combining microarray analysis and gene ontology methods are consistent with previous findings about oxidative DNA damage mechanism. The second publication characterizes EC differentiation process and its impact on EC proliferative capacity in old donor corneas. Analyses of differentiation/progenitor markers and of proliferative capacity underline the differentiation process of EC from the centre to the peripheral corneal endothelium. Thereby, an optimized culture protocol was developed, allowing the formation of high-density monolayer (> 2000 cells/mm2) with stable endothelial morphology. We proved the possibility to make profit from a majority of old-donor cornea grafts invalidated for penetrating graft In the third publication, the activation of endothelial cell cycle by electric pulses directly in corneal graft was characterized. We confirm the activation of endothelial cell cycle at different phases but also the damage of tissue during electroporation. 2- Second strategy consists of the amplification of ECs from potential EC progenitors. Using sphere forming culture and a new method to detect slow-cycling cells, we demonstrate the existence of "young" ECs population with higher proliferative capacity in corneal periphery. The isolation of ECs by sphere formation is one possible step for ECs selection in vitro. 3- The differentiation of embryonic stem cells, mesenchymal stem cells or induced pluripotent stem cells into corneal endothelial cells is the third approach considered by our laboratory. The manipulation of stem cells differentiation would be based on the molecular mechanisms implicated in the formation of corneal endothelium from periocular mesenchymal cells described in the first part of the bibliography. Finally, in order to validate the quality of endothelial cell mass obtained, we revisited recent methods for the evaluation of corneal endothelial identity (immunolocalisation of specific markers), for the measurement of pump activity of cell monolayer (Ussing chamber, perfusion chamber) or directly in deswelled cornea using the bioreactor patented by the BiiGC laboratory
Negoescu, Adrien Nicolas. "Apoptose et évolution phénotypique des cellules corticosurrénaliennes en culture primaire." Grenoble 1, 1995. http://www.theses.fr/1995GRE10010.
Canivet, Ludivine. "Caractérisation et toxicité de nanoparticules manufacturées de fer chez Physcomitrella patens (Hedw. Bruch & Schimp.) et sur cellules épithéliales bronchiques humaines (HBEC) : vers une utilisation en biosurveillance d’aérocontaminants nanoparticulaires." Thesis, Lille 2, 2013. http://www.theses.fr/2013LIL2S042/document.
Many industries emit, unintentionally, ultra-fine particles in the air, for many years. Many questions arise about their effects on ecosystems and human health. In this context, we focused our research on the iron-engineered nanoparticles (Fe-NP), representative of industrial smoke emitted by metallurgical industries and we conducted, in parallel, toxicity and ecotoxicity studies. The main objective of this work was to study the impact of Fe-NP exposed by air in two biological models: Physcomitrella patens (Hedw.) Bruch & Schimp. and primary cultures of human bronchial epithelial cells (HBEC). To meet these objectives, we had to characterize our NP model. Indeed, the physic-chemical characterization must be as complete as possible (8 parameters) to determine, firstly, their surface properties. Then, we checked their penetration within our biological models. And, oxidative stress biomarkers were measured in the bryophyte, exposed to low concentrations of Fe-NP. Firstly, any loss of vitality could be observed over time at the doses tested. Our studies have failed to demonstrate a significant increase of reactive oxygen species and malondialdehyde, and a significant modulation of the ratio GSSG/GSH, although the phenomenon of "over-compensation" can be discussed, over the long term, leading to the production of GSH in our plant showing a plant adaptation to stress. Finally a toxicogenomic analysis showed modulation of expression (not significant) of all isoforms of the genes of interest studied at the doses tested. For toxicity studies, we characterized our cellular model by immunocytological staining. Then, a viability test allowed us to choose the exposure dose: 2 μg.cm-2. The research on oxidative stress and the modulation of gene expression were performed on cells derived from three different patients to take into account individual variability. Unlike some publications, we did not show a dose-dependent increase of ROS. Then, the pangenomic study has allowed us to select 10 genes. A detailed study of this genes showed early effects (from 6 h of exposure) in genes involved in inflammation. However, Fe-NP did not cause any significant increase of MDA and GSSG/GSH ratio after several days of exposure. Following these results, it is now possible to conduct research on the impacts of ultra-fine particles using our two biological models and improve knowledge about their potential danger on the environment and human health
Dias, Gisele Cristiane de Melo. "Caracterização, isolamento e cultura de espermatogônias primárias de curimbatá, Prochilodus lineatus (Valencienes, 1847)." Universidade de São Paulo, 2015. http://www.teses.usp.br/teses/disponiveis/42/42134/tde-08062015-100249/.
Adult males of P. lineatus had their gonads used according routines of light microscopy and transmission electron microscopy. For cell culture, the testes were enzymatically digested; testicular suspension was separated by discontinuous gradient with Percoll followed by adhesion differential plating and the cells were cultured. The enrichment of the spermatogonia was carried by flow cytometry. The testes present three regions with similar distribution of cell types, and nuclear diameter of germ cells decreases significantly during spermatogenesis. The spermatogonia cultured for 15 days with medium for cell proliferation, resulted in large cell agglomerates which were characterized with the antibodies anti-Vasa, anti-GFRa1 and anti-anti-OCT4. The cultures that receiving medium for cell differentiation showed slow proliferation process of primary spermatogonia compared to cell culture medium suggestive for cell differentiation.
String, Andreas Sebastian. "Immunhistochemische Vergleichsanalyse von Primärzellaggregaten und Ursprungsgeweben unterschiedlicher Dignität zur Charakterisirung der in-vitro Anpassung." Doctoral thesis, Humboldt-Universität zu Berlin, Medizinische Fakultät - Universitätsklinikum Charité, 2002. http://dx.doi.org/10.18452/14758.
Immunohistochemical Analysis of primary cell aggregates and their origin tissue of different pathology to evaluate adaptation to in-vitro environment Objective: Three-dimensional cell cultures reflect more closely the in-vitro environment then monolayer cultures. Furthermore, organoid cultures, which contain beside the dominant tumor cell also mesenchymal cells and leucocytes are used to study the interaction of these cells in several aspects of the tumor pathology, such as metastasis, angiogenesis and tumor immunology. Materials and Methods: Specimens obtained from thyroid tissue, thyroid adenomas and carcinomas, ovarian cancer and sarcomas were dissolved to single cell suspensions. After incubation under stirring, primary cell aggregates were cultured within 24-48 hours. Cryostat sections were made and stained with markers of epithelial cells, leucocytes, macrophages, endothelial cells as well as E-cadherin, a2-, a4-, a5- und av Integrin chain, IGF-I und EGF receptor, cerbB2 and Cathepsin D using the APAAP method. The immunhistochemical results of the aggregates and their origin tissue were statistically compared using the Mann-Whitney test. Results: Primary cell aggregates could be obtained from up to 90-100% of all probes. Epithelial cells, leucocytes, macrophages and endothelial cells were found equally in aggregates and their origin tissue. Also E-Cadherin, a4-Integrin, IGF-I und EGF receptors, cerbB2 und Cathepsin D were found equally. The a2-, a5- und av integrin chain was expressed in aggregates of thyroid tissue and adenomas, but not in their origin tissue suggesting a de-novo expression. Conclusion: Primary cell aggregates were easily obtained with the used method and could be used as a model in the study of tumor pathology. The different expressions of integrins show an adaptation to the in-vitro environment and could be a reaction to avoid matrix-related apoptosis.
Triner, Joceline Clare. "Defining neurochemical properties and functions of primary sensory neurons in the rat trigeminal ganglion." Thesis, University of Plymouth, 2013. http://hdl.handle.net/10026.1/1585.
Bonicelli, Jana. "In-vitro-Untersuchungen zu antifibrotischen Wirkungen von β-Adrenozeptoragonisten und Glucocorticoiden in primären equinen Bronchialfibroblasten". Doctoral thesis, Universitätsbibliothek Leipzig, 2015. http://nbn-resolving.de/urn:nbn:de:bsz:15-qucosa-189393.
Silva, William Phillip Pereira da. "Cultura de células osteogênicas primárias a partir de osso de baixa densidade e análise do reparo ósseo periimplantar em ratas osteoporóticas em função da texturização de superfície por meio da oxidação por plasma eletrolítico /." Araçatuba, 2019. http://hdl.handle.net/11449/180686.
Coorientadora: Roberta Okamoto
Banca: Daniela Ponzoni
Banca: Ellen Cristina Gaetti Jardim
Resumo: O objetivo deste estudo foi avaliar um novo método de texturização por PEO com incorporação de Ca e P na superfície do Ti-6Al-4V em ossos de baixa densidade, por meio de avaliação in vitro, ex-in vivo e in vivo, em função de parâmetros topográficos e reparacionais. 57 ratas Wistar (Rattus novergicus), sendo 38 ratas com 6 meses de idade (Grupos OXV - submetidas à ovariectomia e SHAM - cirurgia fictícia) e 19 ratas senis (18 meses de idade: Grupo SENIL), foram divididas para realização do estudo ex-in vivo (n=9) e in vivo (n=48). Os grupos para análise ex-in vivo foram submetidos à eutanásia e os fêmures foram removidos e transportados em meio de cultura contendo meio essencial mínimo modificação alfa (α- MEM) suplementado com 500 µg/mL de gentamicina e 3 µg/mL de fungisona. As células-tronco mesenquimais de medula óssea (CTMs-MO) dos fêmures, foram isoladas e cultivadas em meio de crescimento para manterem-se como CTMs. Após alcançar a subconfluência, as células foram cultivadas em 3 superfícies de discos de Ti-6Al-4V, grupo CONTROLE (superfície usinada) grupo AC (superfície tratada por Ataque Ácido e Jateamento) e grupo PEO (superfície tratada por Oxidação de Plasma Eletrolítico com associação de Cálcio e Fosforo). Para avaliação das respostas celulares foram realizados ensaios de viabilidade celular, expressão gênica de marcadores osteoblásticos, imunolocalização de sialoproteina óssea (BSP) e osteopontina (OPN), atividade da fosfatase alcalina (ALP) e formação de matriz mi... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: The objective of this study was to evaluate a new PEO texturing method with Ca and P incorporation on the Ti-6Al-4V surface in low bone density, by means of in vitro, ex vivo and in vivo evaluation through topographic and repairment parameters. 57 Wistar rats (Rattus novergicus), being 38 at 6 months of age (OXV Groups - submitted to ovariectomy and SHAM surgery) and 19 senile rats (18 months of age: SENIL Group) were divided into three subgroups: ex-in vivo (n = 9) and in vivo (n = 48). The Groups for ex-in vivo analysis were euthanized and femurs were removed and transported in culture medium containing minimal alpha modification (α- MEM) medium supplemented with 500 μg / ml gentamicin and 3 μg / ml fungizone. The mesenchymal stem cells from bone marrow (MSC-M) of the femur were isolated and cultured in growth medium to remain as MSCs. After reaching the subconfluence, the cells were grown on 3 surfaces of Ti-6Al-4V discs, CONTROL group (machined surface) group AC (surface treated by etched-acid) and PEO group (surface treated by Electrolytic Plasma Oxidation with the association of Calcium and Phosphorus). Cell viability assays, gene expression of osteoblastic markers, bone sialoprotein (BSP) and osteopontin (OPN), alkaline phosphatase (ALP) activity, and mineralized matrix formation were performed to evaluate cellular responses. Data were submitted to ANOVA 1 factor test or Kruskal-Wallis test (P <0.05). In the groups for the in vivo study, after 90 days, an implant was i... (Complete abstract click electronic access below)
Mestre
Penna, Vanessa [UNIFESP]. "Alteração da expressão gênica das vias de sinalização TGFβ/BMP, matriz extracelular e moléculas de adesão decorrente da passagem celular em cultura primária de hDPSC". Universidade Federal de São Paulo (UNIFESP), 2014. http://repositorio.unifesp.br/handle/11600/39274.
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Introdução: A Engenharia Tecidual (ET) tem como objetivo a fabricação de órgãos e tecidos. Preconiza-se o uso de células autólogas para evitar a incompatibilidade imunológica, porém, pela escassez do número de células obtidas na fonte celular, muitas passagens se tornam necessárias para atingir um número adequado de células. As culturas primárias freqüentemente sofrem diferenciação indesejada, modificando o comportamento e o destino celular. O uso de enzimas proteolíticas durante as passagens celulares é potencialmente lesivo à fisiologia celular, todavia pouca relevância tem sido dada a este aspecto na ET. Essas enzimas, ao digerir a matriz extracelular (MEC), também alteram proteínas de superfície (PS), interferindo na interação célula-célula (ICC). Consequentemente podem alterar a sinalização celular, a expressão gênica, o comportamento e o destino. Objetivo: Avaliar a expressão gênica na via de sinalização TGFβ/BMP, em matriz extracelular e em moléculas de adesão, das células tronco mesenquimais de polpa dental humana (hDPSC) obtidas diretamente do tecido e sob cultura primária até a terceira passagem. Métodos: Foi analisada a expressão gênica por qRT-PCR array da via de sinalização TGFβ/BMP, matriz extracelular e moléculas de adesão após três passagens celulares na cultura primária de hDPSC. Resultados: Os genes COL16A1(p=0,045), TIMP1(p=0,003), THBS1(p=0,027), TGFBI(p=0,001), ITGA8(p=0,002), FN1(p=0,035), CD44(p=0,046),TSC22D1(p=0,026) e RPL13A(p=0,017) tiveram sua expressão aumentada e os genes NCAM1(p=0,047), BGLAP(p=0,037) e ID1(p=0,027) tiveram sua expressão diminuída em relação às células de tecido de origem. Conclusão: Houve alteração da expressão gênica na cultura celular de hDPSC após três passagens. Porém, um gene solitariamente pode não exercer função chave na diferenciação de células, influenciada pela relação com a MEC e com o ambiente extracelular. O controle por modulação da diferenciação celular representa um aspecto importante no desenvolvimento da ET.
Introduction: Tissue Engineering (TE) aims to manufacture organs and tissues and advocates the use of autologous cells to avoid immune incompatibility. However, a longer culture time is necessary to achieve that purpose. Primary cultures often suffer undesirable differentiation, modifying behavior and cell fate. The use of proteolytic enzymes during cell passages is potentially harmful to cell physiology, but little importance has been given to this aspect in ET. These enzymes which digest the extracellular matrix (ECM) also alter surface proteins (SP) and interfere with cell-cell interactions (ICC). Consequently, may alter cell signaling by modifying gene expression, behavior and cell fate. Objective: To assess gene expression in the signaling TGFβ / BMP pathway, in extracellular matrix and in adhesion molecules, of mesenchymal human dental pulp cells from tissue and cultured until third passage. Methods: We had evaluated gene expression by qRT-PCR array of signaling TGFβ / BMP pathway, in extracellular matrix and in adhesion molecules, of mesenchymal cells from primary and third passage cultured human dental pulp Results: COL16A1(p=0,045), TIMP1(p=0,003), THBS1(p=0,027), TGFBI(p=0,001), ITGA8(p=0,002), FN1(p=0,035), CD44(p=0,046),TSC22D1(p=0,026) and RPL13A(p=0,017) genes had their expression increased and NCAM1(p=0,047), BGLAP(p=0,037) and ID1(p=0,027) genes had reduced expression compared to primary cells. Conclusion: There were modifications in gene expression after three passages. However, a single gene could not be enough to exert a key role in cell differentiation that is broadly affected by its relationship with the ECM and extracellular environment. The control by modulation of cellular differentiation, is an important aspect in the development of ET.
FAPESP: 2013/00288-4
Oster, Sandra. "Untersuchungen zur Dynamik und zum Aggregationsmechanismus von alpha-Synuklein in chronischen Toxinmodellen der dopaminergen Primärzellkultur." Doctoral thesis, Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2018. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-232129.
Mayaudon, Jean-Marie. "Nanostructuration de polymères d'implants neuronaux flexibles, caractérisation de leur biocompatibilité par une étude in-vitro, et conception d'un câble flexible pour l'enregistrement cortical de haute densité." Thesis, Université Grenoble Alpes, 2020. http://www.theses.fr/2020GRALS001.
The research about rehabilitation of motor functions or the exploration of the brain using neuroprostheses has two major issues addressed in this thesis: the implants must be of high biocompatibility in the long term and with a high electrode density. The literature reveals that surface nanostructuring can improve the adhesion and neurite network of neurons and that fine and flexible implants, composed of SU-8 polymers, polyimide or parylene, reduce the glial scar. This thesis shows that a plasma etching (argon, oxygen) forms vertical nanowires (NW) of SU-8 of diameter 50 to 100 nm and long up to 8 μm. NWs can be transposed to an implant made of 10 µm diameter SU8 wires. Also, gold deposition (10 nm) followed by plasma etching leads to vertical NWs of the same diameter and long up to 4,2 µm for polyimide PI-LTC9320, and to shorter "bushy" NWs for parylene C and N, and large diameter 150 nm NWs for parylene HT. The XPS analyzes on the surfaces before and after the plasma have revealed a greater oxygen / carbon ratio while the contact angle (70 to 87°) remain the same or becomes higher. The superhydrophobicity of SU-8 and PI-LTC9320 NWs seems to be explained by the shape of the NWs indicating a Cassie-Baxter configuration. This thesis also shows that 4 µm long SU-8 NWs positively influence the adhesion and neurite network of neurons, compared to 1 µm long SU-8 NWs, for primary cortical cell cultures. However, among the various nanostructured and flat polymers including SU-8, parylenes and polyimides, the flat parylenes reveals to be the best polymers for the cortical cell cultures. In comparison to flat parylene C, nanostructured parylene C has a variable effect on the retinal cell cultures. Also in this thesis, in order to meet the criteria of biocompatibility, sterilization, flexibility and electrical shielding, a flexible cable adapted to the connection of high density soft implants (256 channels) was successfully performed in a clean room (analyzes MEB and EDX). Possible conductive polymers for the shielding have been identified but electrical characterizations remain to be established by connecting the two ends of the cable by a connector developed in this thesis. Finally, the quality of the electrical connection between the cable and the implant, performed easily via a "clip" printed in 3D and anisotropic conductive film has to be improved
Chaves, Gabriela Pena. "Sistema canabinóide e seu possível papel em processos de neuroproteção e plasticidade: estudos in vivo e in vitro." Universidade de São Paulo, 2008. http://www.teses.usp.br/teses/disponiveis/42/42137/tde-03102008-113207/.
The cannabinoid system (CS) seems to have a role in several neurobiological processes, including neuroprotection and neuronal plasticity. The aims of this study were to verify the effects of unilateral retinal ablation on the expression of cannabinoid receptor CB1 and other structural proteins in the optic tectum of chick brain by immunohistochemistry, immunoblotting and real time PCR. Moreover, we evaluated the effects of cannabinoids agonists and antagonists treatment in optic tectum cell cultures exposed to the NMDA by flow cytometry and in the morphology. The retinal ablation seems to generate an increase in the expression of protein CB1 in the deafferented optic tectum, but not in the levels of mRNA. The treatment of the cultures with the cannabinoid agonist decreased the number of unviable cells and fragmented DNAs generated by NMDA. This increase of CB1 expression indicates a post-synaptic localization of these receptors and suggests a role of the CS in plasticity processes. The results of cell culture suggest neuroprotector role of the CS.
Martínez, Romero Carles. "Polycomb group proteins Bmi1 and Ring1B are involved in cell plasticity and tumorigenesis of the pancreas." Doctoral thesis, Universitat Pompeu Fabra, 2009. http://hdl.handle.net/10803/7190.
però no en els acins, en el pàncrees adult. Bmi1 s'induí en cèl·lules acinars durant lesió aguda, en lesions metaplàstiques acinoductals, en neoplàsies intraepitelials pancreàtiques (PanIN) i en PDAC. Ring1B s'incrementà significativament en PanINs de grau alt i en PDAC. La disminució dels nivells de Bmi1 en la línia cel·lular acinar canvià l'expressió dels enzims digestius pancreàtics. Aquests resultats suggereixen que Bmi1 i Ring1B podrien estar contribuint de diferent manera en la progressió tumoral.
Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers. To improve early diagnosis, research efforts are focused in characterising early events of cancer formation like preneoplastic lesions and deciphering the cell origin of the malignancy. Polycomb proteins constitute a family of epigenetic silencers found in a variety of solid tumours. The main hypothesis is that Polycomb might play a role in preneoplastic states in the pancreas and in tumour development and progression. The expression of Bmi1 and RingB was analysed during pancreatic development, in pancreatic tissue from mouse models of disease and in human pancreatic tissue samples. Mechanistic insights of Bmi1 were performed using in vitro models and with induced Bmi1 depletion. Bmi1 and Ring1B were expressed in pancreatic exocrine precursors during early development and in ductal and islet cells, but not in acinar cells, in the adult pancreas. Bmi1 was induced in acinar cells during acute injury, in acinar-ductal metaplastic lesions, in pancreatic intraepithelial neoplasia (PanIN) and PDAC. In contrast, Ring1B was significantly increased in high-grade PanINs and in PDAC. Bmi1 knockdown in acinar cell line changed the expression of pancreatic digestive enzymes. These results suggest that Bmi1 and Ring1B could contribute differently to tumour development.
Moraes, Luis Henrique Rapucci 1983. "Tratamento in vivo e in vitro com a associação de n-ateilcisteina e deferoxamina em camundongos distróficos." [s.n.], 2014. http://repositorio.unicamp.br/jspui/handle/REPOSIP/317503.
Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: Devido ao fato dos camundongos mdx, modelo experimental da distrofia muscular de Duchenne, apresentarem peroxidação lipídica da membrana causada pelo aumento da produção espécies reativas de oxigênio (EROs) no período que antecede o início da degeneração das fibras musculares, sugere-se que o estresse oxidativo pode ser um dos mecanismos primários da degeneração muscular distrófica, ao invés de ser um efeito secundário deste processo. Camundongos mdx tratados com o antioxidante N-acetilcisteína (NAC) apresentaram diminuição da degeneração muscular. De acordo com a literatura a associação de NAC com Deferoxamina (DFX) produz resultado mais efetivo contra o estresse oxidativo do que a administração de NAC isoladamente. Desta forma, o objetivo do presente estudo foi de verificar, através de análises morfológica, celular e bioquímica, se o tratamento in vivo e in vitro com a associação de NAC e DFX diminui a produção das EROs. Para os estudos in vivo foram utilizados camundongos C57BL/10 (grupo controle) e camundongos mdx, com 14 dias de vida pós-natal. Os camundongos mdx e C57BL/10 foram divididos em 4 grupos experimentais: tratados com salina, tratados com NAC+DFX, tratados com DFX e tratados com NAC (150 mg/kg) por 14 dias. Todos os animais foram submetidos à análise de medida de força. Os músculos Esternomastóideo (STN), Diafragma (DIA) e Tibial Anterior (TA) foram retirados e submetidos às técnicas histológicas (HE, azul de Evans e reação de DHE), Western Blotting (TNF-?, NF-?B, MyoD, MAFbx e 4-HNE). Plasma sanguíneo foram utilizadas para determinação de creatina quinase (CK) e de citocinas inflamatórias. Nos experimentos in vitro foram utilizados os músculos do membro pélvico de camundongos C57BL/10 e mdx com 14 dias de vida. As culturas de células musculares foram utilizadas para análises de viabilidade celular (Trypan blue, MTT e vermelho neutro), análise de cálcio e Western Blotting após serem tratadas ou não com NAC e DFX. O tratamento com NAC, DFX e NAC+DFX apresentou efeito benéficos sobre as fibras musculares distróficas, tanto nos experimentos in vivo quanto in vitro, reduzindo a degeneração muscular, a inflamação exacerbada, a peroxidação lipídica e a produção de EROs. Tanto o tratamento isolado dos medicamentos quanto a associação apresentou potencial efeito, entretanto em alguns experimentos a associação mostrou-se mais eficaz contra os danos provocados pela distrofia
Abstract: Due the fact of mdx mice, an experimental model of Duchenne muscular dystrophy, presenting the lipid peroxidation of membrane caused by increased production of reactive oxygen species (ROS) in the period that preced the onset of muscle fibers degeneration, it is suggested that stress oxidative may be one of the primary mechanisms of dystrophic muscle degeneration, rather than a side effect of this process. Mdx mice treated with the antioxidant N-acetylcysteine (NAC) showed a decrease in muscle degeneration. According to the literature the association of NAC with Deferoxamine (DFX) produces more effective results against oxidative stress than NAC alone. Thus, the aim of this study was to verify, through morphological analysis, cellular and biochemistry, if the in vivo and in vitro treatment with the combination of NAC and DFX decreases the ROS production. For in vivo studies were used C57BL/10 mice (control group) and mdx mice, with 14 days postnatal. The mdx and C57BL/10 mice were divided into 4 experimental groups: treated with saline, treated with NAC + DFX, treated with DFX and treated with NAC (150 mg/kg) for 14 days. All animals were subjected to strength measurement analysis. The Sternomastoid (STN), Diaphragm (DIA) and Tibialis anterior (TA) muscles were removed and submitted to histological techniques (HE, Evans blue dye and DHE reaction), Western Blotting (TNF-?, NF-kB, MyoD, MAFbx and 4-HNE). Blood plasma was used for determination of Creatine kinase (CK) and inflammatory cytokines. In the in vitro experiments, the muscles of the pelvic limb of C57BL/10 and mdx mice with 14 days postnatal were used. The muscles culture cells were used for cell viability analysis (Trypan blue, MTT and Neutral red), calcium analysis and Western blotting after being treated or not with NAC and DFX. Treatment with NAC , DFX and NAC + DFX showed benefic effect on dystrophic muscle fibers, both in vivo and in vitro experiments, reducing muscle degeneration, exacerbated inflammation, lipid peroxidation, and ROS production. Either the isolated or the combination treatment of medication showed a potential effect, however in some experiments the combination was more effective against the damage caused by the disease
Doutorado
Anatomia
Doutor em Biologia Celular e Estrutural