Production of apolipoproteins E and A-I by rat hepatocytes in primary culture.

Hepatocytes were isolated from normal adult rat livers and cultured in a modified HI-WO/BA medium. A nearly confluent monolayer was established at the plating concentration employed. The hepatocytes synthesized ansd secreted albumin at rates similar to those observed in vivo. The cells secreted triacylglycerol in the absence of fatty acid substrate. Under these conditions the most abundant triacylglycerol molecular species contained 53 carbons. Incubation with oleic acid markedly increased triacylglycerol secretion predominantly in the form containing a total carbon number of 57. Approx. 80% of the secreted cholesterol was in the free form and this was unaffected by oleic acid. Employing monospecific antibodies constant rates of synthesis and secretion of apolipoproteins E and A-I were demonstrated by quantitative electroimmunoassay of the cell culture media. The rates of albumin, apolipoprotein E, and apolipoprotein A-I production were 1480, 170 and 60 microgram/h per g cell protein, respectively.