Long-term culture of rat hepatocytes on porous membranes in hormonally defined serum-free medium

Abstract

Rat hepatocytes were cultured on type I collagen-coated porous membranes, in Waxman's modified medium supplemented with very low concentrations of insulin (10(-9)m), glucagon (10(-10)m) and dexamethasone (10(-8)m). Under these experimental conditions, specific differentiated functions were well preserved for at least 15 days, as shown by measures of albumin secretion, EROD and PROD activities, by phase contrast microscopy and PAS and ORO staining for intracellular glycogen and lipid contents. These results suggest that this experimental system may be very useful for long-term in vitro pharmacotoxicological studies.