培养人肾远端而非近端小管上皮细胞的转分化。

P C Baer, U W Tunn, G Nunez, J E Scherberich, H Geiger
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引用次数: 34

摘要

人肾近端和远端(厚升肢和早期远端曲小管)上皮细胞已根据其特异性抗原表达分离。流式细胞术、酶细胞化学和电镜对细胞进行了很好的表征,并培养了3个月。培养的小管细胞共表达细胞角蛋白和波形蛋白作为中间丝蛋白。虽然原代离体细胞(近端和远端)显示出其肾元来源的表型特征,但培养的远端细胞显示出去分化/转分化的倾向。远端细胞失去了Tamm-Horsfall糖蛋白的特征表达,并开始重新表达近端标记蛋白氨基肽酶M、γ -谷氨酰转移酶和二肽基肽酶IV。这些抗原在远端细胞中的表达可以通过流式细胞分析和荧光显微镜显示。酶活性测定显示氨肽酶M、γ -谷氨酰转移酶和二肽基肽酶IV有活性,但近端标记酶碱性磷酸酶无活性。这种抗原转移在不同的培养基中都不能被阻止,并且不能恢复原来的表型。通过不同的肽激素对腺苷酸环化酶的激活,培养的细胞显示出其近端和远端来源的特征性激素刺激模式。这些结果表明,考虑到远端标记酶Tamm-Horsfall蛋白的缺失和近端标记酶如二肽基肽酶IV和氨基肽酶M的重新表达,远端小管细胞可能会转分化为更近端的表型。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Transdifferentiation of distal but not proximal tubular epithelial cells from human kidney in culture.

Human renal proximal and distal (thick ascending limb and early distal convoluted tubule) epithelial cells have been isolated according to their specific antigen expression. The cells were well characterized by flow cytometry, enzyme cytochemistry and electron microscopy and cultured for up to 3 months. Cultured tubular cells coexpressed cytokeratin and vimentin as intermediate filament proteins. While primary isolated cells, proximal as well as distal, revealed the phenotypic characteristics of their nephron origin, cultured distal cells showed the tendency to dedifferentiate/transdifferentiate. Distal cells lost their characteristic expression of Tamm-Horsfall glycoprotein and started de novo expression of the proximal marker proteins aminopeptidase M, gamma-glutamyl transferase and dipeptidyl peptidase IV. The expression of these antigens by distal cells could be shown by flow-cytometric analysis and fluorescence microscopy. Enzyme activity assays revealed the activity of aminopeptidase M, gamma-glutamyl transferase and dipeptidyl peptidase IV, but not of the proximal marker enzyme alkaline phosphatase. This antigenic shift could not be prevented in different culture media, and the original phenotype could not be restored. Cultured cells displayed characteristic hormonal stimulation patterns indicative of their proximal and distal origins, as shown by activation of adenylate cyclase by different peptide hormones. These results indicate that distal tubular cells possibly transdifferentiate to a more proximal phenotype in view of loss of the distal marker enzyme Tamm-Horsfall protein and de novo expression of proximal marker enzymes like dipeptidyl peptidase IV and aminopeptidase M.

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