Large-Scale Isolation and Cultivation of Primary Porcine Hepatocytes

Objective To study the cell amount of young pig primary hepatocytes and their functional viability in isolation and culture. Methods The young pig primary hepatocytes were isolated by extracorporeal collagenase perfusion technique. Their viability was assessed by trypan blue exclusion. Hepatocytes were inoculated in the culture medium consisted of 10 % fetal calf serum. The morphological changes of cultured porcine hepatocytes was observed by optical microscope and electronic microscope .The concentrations of albumin and BUN in the supernatant in different culture period were determined. Results The average yield of porcine hepatocytes was 1.55×1010 cells per pig with an average viability of 90.2 %±2.5 %. Hepatocytes had a tendency to aggregate and 85 % of cells formed spheroids within 24 hours in culture and their viability and differentiating function during in vitro culture were valid for 7 days. The viability and differentiating function of hepatocytes were higher in the first three days during cultivation. Conclusion These data indicate that the young pig hepatocytes can be harvested with high yields and retain viability, thus it is an excellent hepatocyte source for bioartificial liver.