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1	Development of new method to enrich human iPSC-derived renal progenitors using cell surface markers / 細胞表面抗原マーカーを用いたヒトiPS細胞由来の腎前駆細胞を濃縮する新規方法の開発 Hoshina, Azusa 25 September 2018 (has links) Supplementary information 追加（2019-09-30) / 京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第21344号 / 医博第4402号 / 新制\|\|医\|\|1031(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授柳田素子, 教授山下潤, 教授江藤浩之 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM iPS cell cell surface marker renal protenitors isolation method cell therapy AKI 490
2	Validation of a novel particle isolation procedure using particle doped tissue samples Patel, J., Lal, S., Wilshaw, Stacy-Paul, Hall, R.M., Tipper, J.L. 01 May 2018 (has links) Yes / A novel particle isolation method for tissue samples was developed and tested using particle-doped peri-articular tissues from ovine cadavers. This enabled sensitivity of the isolation technique to be established by doping tissue samples of 0.25 g with very low particle volumes of 2.5 µm3 per sample. Image analysis was used to verify that the method caused no changes to particle size or morphologies. / The European Union's Seventh Framework Programme (FP7/2007–2013) under grant agreement no. GA-310477, LifeLongJoints. Novel particle isolation method Tissue samples Doping tissue samples Image analysis
3	Mesenchymal stem cell extraction from human umbilical cord tissue : processing to understand and minimise variability in cell yield Iftimia-Mander, Andreea D. January 2013 (has links) Human tissue banks are a potential source of cellular material for the emerging cellbased therapy industry; umbilical cord tissue (UCT) private banking is increasing in such facilities as a source of mesenchymal stem cells for future therapeutic use. However, early handling of UCT is relatively uncontrolled due to the clinical demands of the birth environment and subsequent transport logistics. It is therefore necessary to develop extraction methods that are robust to real world operating conditions,rather than idealised operation. This will be critical for all processes using primary tissue or cell sources. The research work undertaken in this PhD project was initiated by the collaboration with one of the leading private cord blood banks in the UK and later driven by the prospect of expanding the cell therapy and business potential of the bank. The investigation described in this thesis has focused on: - Developing an extraction method for human mesencymal stem cells (hMSCs) from UCT. - Understanding and minimizing the noticed variability in cell yield extracted from UCT by mapping the operating environment and assessing the risk factors before empirically determining their effect on the process. - Establishing the necessary process controls in the production of high quality hMSCs, through a series of wet experiments, targeted at narrowing down the sources of variability down to sub-process level. - Finding a novel method for assessing the cell content and viability of cords prior to processing. Therefore, helping the tissue processing facility to predict the risk of suboptimal cell yield from a given cord tissue section and processing method, given different operating ranges. - Determining the tissue storage requirements and isolation method with acceptable risk of adequate cell recovery. - Characterization of cells extracted from UCT via different extraction methods and comparison to primary cells extracted from other tissue sources. - Investigation of cryopreservation method for UCT. The result of this work provides a solid example of the type of data and analysis that will be required to inform a Quality-by-Design type approach for cell product development and manufacture. It will help tissue processing facilities and banks to predict the probability of cell yields from tissue sections given different operating ranges, and to aid and inform the experimental approach of others. 362.17
4	Etude de l'implication des cellules microgliales et de l'α-synucleine dans la maladie neurodégénérative de Parkinson / Microglia and α-synuclein implication in Parkinson's disease Moussaud, Simon 25 February 2011 (has links) Les maladies neurodégénératives liées à l’âge, telle celle de Parkinson, sont un problème majeur de santé publique. Cependant, la maladie de Parkinson reste incurable et les traitements sont très limités. En effet, les causes de la maladie restent encore mal comprises et la recherche se concentre sur ses mécanismes moléculaires. Dans cette étude, nous nous sommes intéressés à deux phénomènes anormaux se produisant dans la maladie de Parkinson : l’agrégation de l’α-synucléine et l’activation des cellules microgliales. Pour étudier la polymérisation de l’α-synucléine, nous avons établi de nouvelles méthodes permettant la production in vitro de différents types d’oligomères d’α-synucléine. Grâce à des méthodes biophysiques de pointe, nous avons caractérisé ces différents oligomères à l’échelle moléculaire. Puis nous avons étudié leurs effets toxiques sur les neurones. Ensuite, nous nous sommes intéressés à l’activation des microglies et en particulier à leurs canaux potassiques et aux changements liés au vieillissement. Nous avons identifié les canaux Kv1.3 et Kir2.1 et montré qu’ils étaient impliqués dans l’activation des microglies. En parallèle, nous avons établi une méthode originale qui permet l’isolation et la culture de microglies primaires issues de cerveaux adultes. En comparaison à celles de nouveaux-nés, les microglies adultes montrent des différences subtiles mais cruciales qui soutiennent l’hypothèse de changements liés au vieillissement. Globalement, nos résultats suggèrent qu’il est possible de développer de nouvelles approches thérapeutiques contre la maladie de Parkinson en modulant l’action des microglies ou en bloquant l’oligomérisation de l’ α-synucléine. / Age-related neurodegenerative disorders like Parkinson’s disease take an enormous toll on individuals and on society. Despite extensive efforts, Parkinson’s disease remains incurable and only very limited treatments exist. Indeed, Parkinson’s pathogenesis is still not clear and research on its molecular mechanisms is ongoing. In this study, we focused our interest on two abnormal events occurring in Parkinson’s patients, namely α-synuclein aggregation and microglial activation. We first investigated α-synuclein and its abnormal polymerisation. For this purpose, we developed novel methods, which allowed the in vitro production of different types of α-synuclein oligomers. Using highly sensitive biophysical methods, we characterised these different oligomers at a single-particle level. Then, we tested their biological effects on neurons. Afterwards, we studied microglial activation. We concentrated our efforts on two axes, namely age-related changes in microglial function and K+ channels in microglia. We showed that Kv1.3 and Kir2.1 K+ channels are involved in microglial activation. In parallel, we developed a new approach, which allows the effective isolation and culture of primary microglia from adult mouse brains. Adult primary microglia presented subtle but crucial differences in comparison to microglia from neo-natal mice, confirming the hypothesis of age-related changes of microglia. Taken together, our results support the hypotheses that microglial modulation or inhibition of α-synuclein oligomerisation are possible therapeutic strategies against Parkinson's disease. Vieillissement Maladies neurodégénératives Maladie de Parkinson Neurodégénération Α-synucléine Oligomères Toxicité Dopamine Neurones Neuroinflammation Microglies Immunité du cerveau Lignée cellulaire C8-B4 Cultures primaires Méthode d’isolation in vitro Patch-clamp Electrophysiologie Canaux potassiques Kv1.3 - Kir2.1 Oxyde nitrique Cytokines Aging Neurodegenerative disorders Parkinson’s disease Neurodegeneration Α-synuclein Oligomers Toxicity Dopamine Neurons Neuroinflammation Microglia Brain macrophages Brain immunity C8-B4 cell line Primary culture In vitro isolation method Patch-clamp Electrophysiology Potassium channels Kv1.3 - Kir2.1 Nitric oxide Cytokines 612 616.9

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