AJP - Cell Physiology, Vol 251, Issue 5 780-C786, Copyright © 1986 by American Physiological Society
Use of monoclonal antibodies to culture rat proximal tubule cells
R. C. Stanton, D. L. Mendrick, H. G. Rennke and J. L. Seifter
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.
