Isolation of hepatocytes from postnatal mice.

A method for the isolation of hepatocytes from postnatal (1- to 3-week-old) mice has been developed. Cell isolation was carried out by retrograde perfusion of the liver with a collagenase-containing bicarbonate buffer. Viable cells were separated by selective adsorption onto collagen membranes. Cell viability was assessed by measuring ATP and glutathione content, lactate:pyruvate ratio, and the rate of protein synthesis. Comparisons of these parameters were made with those in cells isolated from adult mice and with values in whole liver. These hepatocytes were capable of metabolizing acetaminophen to its known hepatic conjugates and were susceptible to acetaminophen toxicity. This procedure for isolating mouse hepatocytes should be useful for the study of hepatic drug metabolism and its relationship to toxicity in the postnatal mouse.