The effect of collagenase and duration of liver perfusion on isolation of rat hepatocytes

Objective To study the method of hepatocyte isolation.Methods Using two defferent collagenases (0.05%,type Ⅰ and Ⅳ) the rat hepatocytes were isolated. The effects of defferent types of collagenases and different duration of liver perfusion(5,10 and 15 min) on viability and outputs of hepatocytes were investigated and the effect of culture on viability in 0.02% collagenase solution studied. Results The best results of isolation were accessed by collagenase typeⅣ.Cell viability of two groups(Ⅳ and Ⅰ) was (89.5±3.5)% and (58.0±14.0)%,output (2.5±0.2)×10 11 cell/L and (0.9±0.5)×10 11 cell/L following 10 min perfusion (P0.01) respectively. The structure and functions were normal by culture and biochemical assay.The study also showed that cell viability was increased by the method of cultue in low concentration of collagenase solution.Conclusion Collagenase type Ⅳ and 10 min liver perfusion were the best conditions for rat hepatcyte isolation.