Preparation of rat liver cells: I. Effect of Ca 2+ on enzymatic dispersion of isolated, perfused liver

Abstract

Suspensions of rat liver cells were prepared by perfusion of the isolated liver with collagenase and hyaluronidase. By means of a novel dispersion assay, which measured swelling of the liver in a closed perfusion system, the time course of enzymatic dispersion could be followed. Ca 2+ stimulated the enzymatic dispersion strongly, but a preliminary removal of Ca 2+ with the chelator EGTA rendered the liver tissue more susceptible to the action of enzymes. The best result was thus obtained when the liver was first perfused 5 min with EGTA, then 5 min with enzymes and Ca 2+. This sequential treatment converted the whole liver to a cellular suspension, in which about 95% of the cells were intact.