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Mechanism of regeneration of renal tubular cells and differentiation from ES and iPS cells to renal tubular cells

肾小管细胞再生及ES、iPS细胞向肾小管细胞分化的机制

基本信息

批准号：
21591038
负责人：
MONKAWA Toshiaki
金额：
$ 2.91万
依托单位：
Keio University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2009
资助国家：
日本
起止时间：
2009 至 2011
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21591038/
关键词：
腎臓尿細管再生医学 ES細胞 iPS細胞 ips細胞

项目摘要

The S3 segment of the kidney proximal tubule is highly susceptible to ischemia insults but has a remarkable capacity to repair its structure and function. By applying the "toxin receptor mediated cell knockout" method under the control of the S3 segment-specific promoter Gsl5, we established a transgenic mouse line expressing the human diphtheria toxin(DT) receptor only in the S3 segment. The administration of DT to these transgenic mice caused the selective ablation of S3 segment cells in a dose-dependent manner. These results indicate that this transgenic mouse can suffer acute kidney injury(AKI) caused by S3 segment-specific damage after DT administration. This transgenic line offers an excellent model to uncover the mechanisms of AKI and its rapid recovery.We tried to establish differentiation method of ES cells and iPS cells into renal tubular cells. In ES cells, Activin enhanced the differentiation of ES cells to tubular cells, judging by expression of Ksp-Cadherin, the epithelial marker. Activin also enhanced the differentiation of iPS cells to tubular cells, although the enhancement was lower than in ES cells. We generated monoclonal antibody against the extracellular domain of Ksp-Cadherin. Flow cytometry using the antibody purified the Ksp-positive cells, which formed tubular structure on Matrigel. We established the differentiation method of ES and iPS cells into renal tubular cells.

肾近端小管S3段对缺血损伤高度敏感，但具有显著的修复其结构和功能的能力。通过应用S3片段特异性启动子Gsl 5控制下的“毒素受体介导的细胞敲除”方法，我们建立了仅在S3片段中表达人白喉毒素（DT）受体的转基因小鼠系。DT对这些转基因小鼠的给药以剂量依赖的方式引起S3节段细胞的选择性消融。这些结果表明，该转基因小鼠在DT给药后可遭受由S3片段特异性损伤引起的急性肾损伤（阿基）。本研究尝试建立ES细胞和iPS细胞向肾小管细胞分化的方法。在ES细胞中，通过上皮标记物Ksp-钙粘蛋白的表达来判断，激活素增强ES细胞向肾小管细胞的分化。激活素也促进iPS细胞向肾小管细胞的分化，尽管这种促进作用低于ES细胞。我们制备了抗Ksp-Cadherin胞外区的单克隆抗体。使用抗体的流式细胞术纯化Ksp阳性细胞，其在Matrigel上形成管状结构。建立了ES细胞和iPS细胞向肾小管细胞分化的方法。