Use of monoclonal antibodies to culture rat proximal tubule cells.
Current renal cell culture techniques are limited by either a low yield of cells or by heterogeneity of cell types. We have used monoclonal antibodies to microvillus membrane proteins to isolate and culture a pure population of proximal tubule cells. The cells were characterized as proximal tubule cells by phase microscopy, enzyme histochemistry for alkaline phosphatase, butyrate esterase, and gamma-glutamyltransferase, electron microscopy, and specific reactivity with a variety of monoclonal antibodies for proximal tubule cells. Growth over 2-7 days yielded cell numbers up to 1,000-fold greater than obtained by single tubule microdissection. Dome formation was observed, suggesting intact fluid transport. In addition, Na+-H+ exchange and Na+-dependent D-hexose transport, known transport processes of the proximal tubule, were demonstrated by microfluorimetry of single cells and methyl-alpha-D-glucopyranoside uptake, respectively. Our results indicate that large numbers of homogeneous, cultured rat proximal tubule cells that maintain characteristics of in vivo proximal tubule cells can be obtained using a monoclonal antibody technique of isolation.
D000818:Animals; D000911:Antibodies, Monoclonal; D000941:Antigens / Q000276:immunology; D002352:Carrier Proteins / Q000378:metabolism; D002455:Cell Division; D002469:Cell Separation / Q000379:methods; D006651:Histocytochemistry; D007687:Kidney Tubules, Proximal / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D008871:Microvilli / Q000276:immunology; D051381:Rats; D011919:Rats, Inbred Strains; D017923:Sodium-Hydrogen Exchangers
D000911:Antibodies, Monoclonal; D000941:Antigens; D002352:Carrier Proteins; D017923:Sodium-Hydrogen Exchangers