Incubation of HK and LK sheep red cells in vitro for long periods

Abstract

HK and LK sheep red cells have been maintained in vitro at 37°C for periods of up to 3 weeks. During the incubation, periodic measurements of cell Na +, K +, ouabain-sensitive and ouabain-insensitive K + influxes were made. The effects of lowering the in vitro incubation temperature and of reducing the K + concentration in the medium on cell cation content and volume were also investigated. The results of these experiments were, in general, consistent with previous measurements of the effects of these parameters on K + and Na + fluxes in short term experiments. In vitro incubation for several days was also used to show that exposure to anti-L serum produces a net uptake of K + and extrusion of Na + in LK sheep red cells. A new procedure for measuring the active and passive influxes of K + was developed. The early time course of K + influx inhibition by ouabain was also studied by this procedure.