Isolation, propagation and characterization of primary tubule cell culture from human kidney
Isolation, propagation and characterization of primary tubule cell culture from human kidney
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DOI：
10.1111/j.1440-1797.2007.00779.x
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发表时间：
2007-04-01
期刊：
影响因子：
2.5
通讯作者：
Chen, Xinming
中科院分区：
文献类型：
--
作者：
Qi, Weier;Johnson, David W.;Chen, Xinming
近端小管细胞（PTC）是皮质小管间质中的主要细胞类型。由于PTC在肾小管间质病理生理学中发挥着核心作用，因此从肾组织中制备纯PTC对于探索肾小管间质病理机制至关重要。作者使用 Percoll 密度梯度离心作为关键的 PTC 富集步骤，成功地精炼并表征了人类 PTC 的原代培养物。通过这种方法获得的细胞保留了 PTC 的形态和功能特性，并且被其他肾细胞的污染降至最低。特别是，初级分离株具有上皮细胞的特征，具有均匀的极化形态、紧密连接和形态良好的顶端微绒毛。细胞角蛋白在分离株中均匀且强烈表达。刷状缘酶活性和 PTC 转运特性保留在分离株中。因此，该方法为人类近曲小管的生理研究提供了一个极好的体外模型。
Proximal tubule cells (PTC) are the major cell type in the cortical tubulointerstitium. Because PTC play a central role in tubulointerstitial pathophysiology, it is essential to prepare pure PTC from kidney tissue to explore the mechanisms of tubulointerstitial pathology. The authors have successfully refined and characterized primary cultures of human PTC using Percoll density gradient centrifugation as a key PTC enrichment step. The cells obtained by this method retain morphological and functional properties of PTC and are minimally contaminated by other renal cells. In particular, the primary isolates have characteristics of epithelial cells with uniform polarized morphology, tight junction and well-formed apical microvilli. Cytokeratin is uniformly and strongly expressed in the isolates. Brush border enzyme activities and PTC transport properties are retained in the isolates. This method therefore provides an excellent in vitro model for the physiologic study of the human proximal tubule.