LACK OF EFFECT OF EXTRACELLULAR-MATRIX OR 3T3 FEEDER LAYER ON THE MAINTENANCE OF DIFFERENTIATION OR SURVIVAL-TIME OF CULTURED RAT HEPATOCYTES

Cultured rat hepatocytes are often used in in vitro studies. In this c ulture system the lack of the normal environment to which the cells wo uld be exposed in vivo contributes to a change in differentiation. To study this differentiation problem we compared the effects of culturin g primary hepatocytes on an extracellular matrix (ECM) isolated from n ormal rat livers and on a 3T3 (mouse fibroblast) feeder layer. The use of the two coatings did not result in changes in survival time, as me asured by leakage of lactate dehydrogenase, differentiation (activitie s of pyruvate kinase and gamma-glutamyltransferase) or specific functi ons (albumin production and concentration of cytochrome P-450) in comp arison with cells cultured on plastic. Since no positive effects of th e coatings were found it is proposed that more attention should be pai d to methods of isolating and culturing hepatocytes, rather than to ob taining conditions for the hepatocytes that are completely comparable with the in vivo situation.