Blais A, Jalal F, Crine P, Paiement J, Berteloot A
Department of Physiology, Faculty of Medicine, University of Montreal, Quebec, Canada.
Am J Physiol. 1992 Jul;263(1 Pt 2):F152-62. doi: 10.1152/ajprenal.1992.263.1.F152.
We have determined the influence of glucose (Glc)-free medium on the growth and differentiation of rabbit kidney proximal tubule cells (PTC) in primary cultures. The specific growth rates and the protein-to-volume ratios are shown to be independent of the culture conditions. In contrast, the functional expression of four brush-border membrane enzyme markers was found to decline steadily and in the same way from day 3 in culture up to late confluence in Glc-containing medium, and different evolution patterns and high expression levels were observed up to confluence in a Glc-free glutamine (Gln)-supplemented medium. Electron microscopy clearly showed, however, that the functional and morphological differentiation of the brush-border membrane is not correlated. Finally, by use of an indirect immunofluorescent technique in combination with flow cytometry, it is demonstrated that confluent cells grown in Glc and Gln media form homogeneous cell populations of PTC. It is thus concluded that the functional differentiation of rabbit kidney PTC in primary cultures is strongly dependent upon the energy source present in the culture medium.
我们已经确定了无葡萄糖（Glc）培养基对原代培养的兔肾近端小管细胞（PTC）生长和分化的影响。结果显示，比生长速率和蛋白质与体积比与培养条件无关。相反，在含葡萄糖的培养基中，从培养第3天到汇合后期，四种刷状缘膜酶标志物的功能表达呈稳定且相同方式下降；而在添加无葡萄糖谷氨酰胺（Gln）的培养基中，直至汇合时观察到不同的演变模式和高表达水平。然而，电子显微镜清楚地表明，刷状缘膜的功能和形态分化并无相关性。最后，通过使用间接免疫荧光技术结合流式细胞术，证明在葡萄糖和谷氨酰胺培养基中生长的汇合细胞形成了PTC的同质细胞群。因此得出结论，原代培养的兔肾PTC的功能分化强烈依赖于培养基中存在的能量来源。
