[Free flow electrophoresis. Application to the separation of 2 populations of proximal tubule cells from the rabbit kidney]

Abstract

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.