Study on the isolation, primary culture and biologicial functions of rat hepatocytes

Objectives:To investigate the methods of isolation of rat hepatocytes and observe the biologicial characteristic of primary culture rat hepatocytes. Methods:Hepatocytes were isolated from SD rat by collagenase digestion and cultured in a conditioned medium. The viability were assessed by trypan blue exclusion. The morphologic change of cultured hepatocytes was observed, the concentrations of albumin, urea and lactate dehydrogenase(LDH) in the supernatant in different cultural period were examined. Results: Hepatocytes obtained by collagenase digestion were intact and have a good viability and function. The LDH leakage, albumin synthesis and urea level fluctuate in one week,there is a lesser LDH leakage and higher albumin and urea level on the third day. Conclusions: The collagenase digestion method is feasible for isolation of hepatocytes. The primary culture rat hepatocytes have a good function in the third day.