Establishment of an improved collagenase perfusion technique to isolate the hepatocyte effectively

Objective To establish an efficient, effective hepatocyte isolation technique in order to increase cell production and decrease the prime cost.Methods An improved collagenase perfusion isolation technique was established to isolate mouse hepatocytes by a two-step collagenase perfusion in situ.Hepatocytes were purified by centrifugation with Percoll, the hepatocytes yields and viabilities were measured.The difference of hepatocyte yields and viability were compared between the collagen slicing digestion group and the improved technique group.Results We establish an improved collagenase perfusion isolation technique.The isolation protocol used result in a high cell yield of (1.74±0.35) ×107/L and the viability is 90.1%±1.9%.The yield of isolated hepatocyte using collagenase slicing digestion technique is (0.44±0.06) ×107/L and the viability is 35.3%±1.4%, P<0.01.Conclusion The improved mouse hepatocyte isolation technique is simple and easy, the yield and viability of isolated hepatocyte is high.