Toutain H, Morin J P, Fillastre J P
INSERM U 295, UER de Médecine de Rouen, Saint-Etienne-du-Rouvray, France.
Pathol Biol (Paris). 1989 Oct;37(8):869-74.
Two cell populations from the proximal tubule of the rabbit kidney were separated by free flow electrophoresis from a pure suspension of proximal tubular cells obtained by a combination of a Ca-binding agent, gentle mechanical forces and differential sifting. Before the electrophoretic separation, distal and proximal enzyme activities were measured on the cortical homogenates, on the proximal tubule suspensions and on the isolated cell samples in order to assess the purity of the cell preparation. The isolated cells were very poor in distal tubule marker activities and were enriched in proximal tubule marker enzymes. Cell oxygen consumption was measured before and after the electrophoretic run were similar and reflected high cell metabolic capacity. The cells in the slow-moving electrophoresis fractions had a high gamma-glutamyl transpeptidase activity and the fast moving cells showed a high glucose-6-phosphatase activity. These results point out a separation of viable cells from straight and convoluted portion of the proximal tubule from the rabbit kidney. These two cell populations can be suitable for further use in biochemical and physiological studies.
通过自由流动电泳从兔肾近端小管的纯细胞悬液中分离出两个细胞群体,该细胞悬液是通过结合使用钙结合剂、轻柔的机械力和差异筛分获得的。在进行电泳分离之前,对皮质匀浆、近端小管悬液和分离出的细胞样品进行远端和近端酶活性测量,以评估细胞制备物的纯度。分离出的细胞中远端小管标记物活性很低,而近端小管标记酶则很丰富。在电泳运行前后测量细胞耗氧量,结果相似,反映出细胞具有较高的代谢能力。电泳迁移较慢部分的细胞具有较高的γ-谷氨酰转肽酶活性,而迁移较快的细胞则具有较高的葡萄糖-6-磷酸酶活性。这些结果表明从兔肾近端小管的直部和曲部分离出了活细胞。这两个细胞群体可适合进一步用于生化和生理学研究。