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Establishment of an in vitro co-culture system for the renal glomerular filtration barier.

肾小球滤过屏障体外共培养系统的建立。

基本信息

批准号：
14570015
负责人：
KOBAYASHI Naoto
金额：
$ 2.5万
依托单位：
Ehime University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570015/
关键词：
kidney glomerulus glomerular filtration barrier podocyte endothelial cell cell culture cytoskeleton cell adhesion

项目摘要

The initial step of the urine generation in the kidney is the glomerular filtration through the glomerular filtration barrier, which prevents protein leakage into the urine. In order study the pathophysiology of glomerular diseases, where the ultrastructure of the glomerular filtration barrier is damaged, it is necessary to establish a reliable model system to reproduce the structure and function of the barrier. The filtration barrier is composed of three components, i.e. the glomerular capillary endothelial cell, the glomerular basement membrane, and the podocyte, also known as the glomerular epitheliaI cell. This study has aimed to establish such a model using cell culture system.With the conditionally immortalized mouse podocyte cell line which is kindly provided by our foreign collaborators, we have studied the signal transduction pathway regulating the podocyte morphogenesis (both process formation and cell-cell contact formation) with special reference on the RHO-family small GTP … More ases (RhoA, Rac1, Cdc42) (Kobayashi N, 2002 ; Kobayashi et al., 2004 ; Gao et al., 2004 ; Gao et al., manuscript in preparation). Since in in vivo glomeruli, podocytes develop a cell-type specific intercellular junction, slit membrane, the formation of intercellular junctions between podocytes is thought to be the critical step in the development of the filtration barrier. We have shown that tight cell-cell contacts are induced between cultured podocytes by inhibition of RhoA with forskolin. Biochemical approach revealed the activation of other members of the RHO-family, Rac1 and Cdc42, during this cell-cell contact formation. It seems unlikely that a co-culture system (podocytes + endothelial cells) is absolutely necessary to establish the glomerular filtration barrier in vitro. We have also shown that pharmacological inhibition of RhoA and its downstream effector, ROCK, enhances process formation of the cultured podocytes. As planned at the beginning of this study, we are still trying to establish cell lines of renal glomerular endothelial cells. Less

肾脏中尿液生成的初始步骤是通过肾小球过滤屏障的肾小球过滤，其防止蛋白质渗漏到尿液中。为了研究肾小球疾病的病理生理学，其中肾小球滤过屏障的超微结构被破坏，有必要建立一个可靠的模型系统来再现屏障的结构和功能。滤过屏障由三种成分组成，即肾小球毛细血管内皮细胞、肾小球基底膜和足细胞（也称为肾小球上皮细胞）。本研究利用细胞培养系统建立足细胞形态发生模型，利用国外合作者提供的条件永生化小鼠足细胞系，研究了调节足细胞形态发生（突起形成和细胞间接触形成）的信号转导途径，并以RHO家族小GTP为研究对象，探讨了其在足细胞形态发生中的作用 ...更多信息酶（RhoA，Rac 1，Cdc 42）（小林N，2002 ;小林等人，2004 ; Gao等人，2004年; Gao等人，手稿正在准备中）。由于在体内肾小球中，足细胞形成细胞类型特异性的细胞间连接，狭缝膜，足细胞之间细胞间连接的形成被认为是形成滤过屏障的关键步骤。我们已经表明，通过用毛喉素抑制RhoA，在培养的足细胞之间诱导紧密的细胞-细胞接触。生物化学方法揭示了在这种细胞-细胞接触形成过程中RHO-家族的其他成员Rac 1和Cdc 42的激活。共培养系统（足细胞+内皮细胞）似乎不太可能是体外建立肾小球滤过屏障所必需的。我们还表明，药理学抑制RhoA及其下游效应，ROCK，增强过程的形成培养的足细胞。正如本研究开始时的计划，我们仍在尝试建立肾小球内皮细胞的细胞系。少