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 culture system the lack of the normal environment to which the cells would be exposed in vivo contributes to a change in differentiation. To study this differentiation problem we compared the effects of culturing primary hepatocytes on an extracellular matrix (ECM) isolated from normal 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 measured by leakage of lactate dehydrogenase, differentiation (activities of pyruvate kinase and gamma-glutamyltransferase) or specific functions (albumin production and concentration of cytochrome P-450) in comparison with cells cultured on plastic. Since no positive effects of the coatings were found it is proposed that more attention should be paid to methods of isolating and culturing hepatocytes, rather than to obtaining conditions for the hepatocytes that are completely comparable with the in vivo situation.