Differentiation status of human renal proximal and distal tubular epithelial cells in vitro: Differential expression of characteristic markers
Differentiation status of human renal proximal and distal tubular epithelial cells in vitro: Differential expression of characteristic markers
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DOI：
10.1159/000096947
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发表时间：
2006-01-01
影响因子：
2.7
通讯作者：
Geiger, Helmut
中科院分区：
文献类型：
--
作者：
Baer, Patrick C.;Bereiter-Hahn, Juergen;Geiger, Helmut
背景：肾元来源明确的人肾小管细胞的培养是研究肾脏各种生理和病理生理机制的重要基础。体外培养细胞的分化受细胞分离和培养条件的影响。我们的研究描述了体外培养的人肾近端和远端（厚升肢和早期远端）小管上皮细胞的分化。方法：采用免疫磁法分离近端小管细胞（PTC）和早期远端小管细胞（DTC）并进行培养。体外分化采用Western blot分析和特征标记逆转录聚合酶链反应进行评估。荧光和扫描电镜显示了其形态特征。结果：E-cadherin在DTC中高表达，在PTC中低表达。与DTC相比，在PTC中，细胞间粘附分子1和水通道蛋白1组成性表达水平较高。两种细胞类型均表达细胞角蛋白18和na - k - atp酶，但不表达α -平滑肌肌动蛋白。形态学分析表明，PTC的微绒毛较长，DTC的微绒毛较短，两者均表达紧密连接蛋白闭塞带蛋白1。结论：在体外培养的PTC和DTC中，E-cadherin、细胞间粘附分子1和水通道蛋白1的表达存在差异。我们可以显示细胞的体外分化，因此，它们作为人类近端和早期远端小管的体外模型的实用性。版权所有(c) 2006 S. Karger AG，巴塞尔。
Background: The culture of human renal tubular cells of well-defined nephron origin is an important basis in the research of various physiological and pathophysiological mechanisms in the kidney. In vitro differentiation of cultured cells is affected by cell isolation and culture conditions. Our study describes in vitro differentiation of cultured human renal proximal and distal (thick ascending limb and early distal) tubular epithelial cells. Methods: Proximal tubular cells (PTC) and early distal tubular cells (DTC) were isolated immunomagnetically and cultured. In vitro differentiation was assessed by Western blot analysis and reverse transcriptase polymerase chain reaction using characteristic markers. Morphologic characterization was shown by fluorescence and scanning electron microscopy. Results: E-cadherin was highly expressed in DTC, whereas expression was low in PTC. In contrast to DTC, in PTC, intercellular adhesion molecule 1 and aquaporin 1 were constitutively expressed at high levels. Both cell types expressed cytokeratin 18 and Na-K-ATPase but not alpha-smooth muscle actin. Morphological analysis showed that PTC develop long microvilli, whereas DTC formed short microvilli in culture, and both express the tight junction protein zona occludens protein 1. Conclusions: We demonstrate differential expression of E-cadherin, intercellular adhesion molecule 1 and aquaporin 1 in cultured PTC and DTC as described for the renal tubule in vivo. We could show the in vitro differentiation of the cells, and thus, their usefulness as an in vitro model of the human proximal and early distal tubule. Copyright (c) 2006 S. Karger AG, Basel.