Abstract: Objective To investigate the method of isolation and extraction of tilapia hepatocytes. Methods The fish liver was washing repeatedly by D-Hank, then the liver was cut into small pieces. The tissues was filtered and centrifuged after digested by Collagenase IV. Then the suspensions were purified by Percoll isolation medium after washed by DMEM. Hapatocytes was counted. The morphologic characteristics of the isolated hapatocytes were accessed and observed by light microscope. Result An amount mixed cells were found through the cold digestion of Collagenase method. Further isolation was done by centrifuged at 90 g, 60 g and 30g. Conclusion The method of Cold digestion by Collagenase had obtained the best isolated result. Better purification and amount of hepatocytes could be done by centrifugation with 60g for 2 minutes and centrifugation with 30g for 3 minutes.

Key words: Collagenase, Cold digestion, Hepatocytes, Tilapia, Isolation