Perfusion technique of suckling rat liver, and comparison of cytologic and biochemical properties between hepatocytes isolated from suckling and adult rats.

Collagenase-liver-perfusion technique, currently used with adult rat liver, was applied to isolation of hepatocytes from suckling rat liver. The total hepatocyte numbers isolated from suckling rats by collagenase-liver-perfusion technique were 9-fold higher than those by non-perfusion technique. The yield and viability of isolated hepatocytes from suckling rats were 18.1 X 10(7) cells per gram liver and 95%, respectively. The cell yield per gram liver and viability from suckling rats were 185% and 112% of those from adult ones, respectively. In comparison with adult rat hepatocytes, suckling rat hepatocytes were smaller and more homogeneous. The percentage (3.1%) of binucleate cells in the hepatocytes isolated from suckling rats was about one tenth of that (30.7%) in the hepatocytes isolated from adult rats. The isolated suckling rat hepatocytes had higher tyrosine aminotransferase (TAT) and glucose-6-phosphate (G6Pase) activities than adult ones. Suckling and adult rat hepatocytes were transferred to primary culture to compare their cell number kinetics and functional longevities. The functional longevities of those hepatocytes were assessed by their capacity to secrete albumin and alpha-fetoprotein (AFP) into the culture medium and to express TAT and G6Pase activities up to day 6 of primary culture.