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培养中的足细胞:过去、现在与未来。

Podocytes in culture: past, present, and future.

作者信息

Shankland S J, Pippin J W, Reiser J, Mundel P

机构信息

Division of Nephrology, Department of Medicine, University of Washington, Seattle, Washington, USA.

出版信息

Kidney Int. 2007 Jul;72(1):26-36. doi: 10.1038/sj.ki.5002291. Epub 2007 Apr 25.

Abstract

Human genetic and in vivo animal studies have helped to define the critical importance of podocytes for kidney function in health and disease. However, as in any other research area, by default these approaches do not allow for mechanistic studies. Such mechanistic studies require the availability of cells grown ex vivo (i.e., in culture) with the ability to directly study mechanistic events and control the environment such that specific hypotheses can be tested. A seminal breakthrough came about a decade ago with the documentation of differentiation in culture of primary rat and human podocytes and the subsequent development of conditionally immortalized differentiated podocyte cell lines that allow deciphering the decisive steps of differentiation and function of 'in vivo' podocytes. Although this paper is not intended to provide a comprehensive review of podocyte biology, nor their role in proteinuric renal diseases or progressive glomerulosclerosis, it will focus specifically on several aspects of podocytes in culture. In particular, we will discuss the scientific and research rationale and need for cultured podocytes, how podocyte cell-culture evolved, and how cultured podocytes are currently being used to uncover novel functions of podocytes that can then be validated in vivo in animal or human studies. In addition, we provide a detailed description of how to properly culture and characterize podocytes to avoid potential pitfalls.

摘要

人类遗传学研究和体内动物研究有助于明确足细胞在健康和疾病状态下对肾功能的关键重要性。然而,与其他任何研究领域一样,默认情况下这些方法无法进行机制研究。此类机制研究需要利用体外培养的细胞(即在培养皿中培养),以便能够直接研究机制性事件并控制环境,从而检验特定假设。大约十年前取得了一项重大突破,记录了原代大鼠和人类足细胞在培养中的分化情况,随后开发出条件永生化分化足细胞系,这使得人们能够解读“体内”足细胞分化和功能的决定性步骤。尽管本文无意全面综述足细胞生物学,也不涉及它们在蛋白尿性肾病或进行性肾小球硬化中的作用,但将特别关注培养中的足细胞的几个方面。具体而言,我们将讨论培养足细胞的科学和研究原理及必要性、足细胞培养的发展历程,以及目前如何利用培养的足细胞来揭示足细胞新功能,进而在动物或人体研究中进行体内验证。此外,我们还详细描述了如何正确培养和鉴定足细胞以避免潜在问题。

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