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.

肾脏近端小管的S3段非常容易受到缺血损伤的影响，但具有显着的修复其结构和功能的能力。通过将“毒素受体介导的细胞敲除”方法应用于S3段特异性启动子GSL5的控制，我们建立了仅在S3段中表达人二骨毒素（DT）受体的转基因小鼠系。这些转基因小鼠的DT给药以剂量依赖性方式导致S3节段细胞的选择性消融。这些结果表明，这种转基因小鼠可能遭受由DT给药后S3段特定损伤引起的急性肾损伤（AKI）。该转基因线提供了一个出色的模型，可揭示AKI的机制及其快速恢复。我们试图将ES细胞和IPS细胞的分化方法建立到肾小管细胞中。在ES细胞中，激活素增强了ES细胞对管状细胞的分化，通过表达上皮标记的KSP-钙粘着蛋白的表达来判断。激活素还增强了IPS细胞对管状细胞的分化，尽管增强量低于ES细胞。我们产生了针对KSP-钙粘蛋白细胞外结构域的单克隆抗体。使用抗体纯化KSP阳性细胞的流式细胞仪，该细胞在基质凝胶上形成管状结构。我们将ES和IPS细胞的分化方法建立到肾小管细胞中。