Isolation, cuture and identification of hepatic non-parenchymal cells in rats

Objective To establish an effective method to isolate hepatic non-parenchymal cells including hepatic stellate cells (HSCs),sinusoidal endothelial cells (SECs) and macrphages respectivly.Methods The whole livers were removed from rats under sterile condition,then the livers were perfused with digestive enzyme in vitro and cut into species.The species were digested to generate single cells suspension at 37 ℃.By using the methords of Percoll gradient centrifugation and adherent culture,the single cells were isolated into HSCs,SECs,and macrophages.The cell activity was observed by the trypan blue excluding test,and the cell purity was identified using spontaneous fluorescence,immunofluorescence and cellular morphology.Results The purity of hepatic non-parenchymal cells was more than 90％,and the survival rate of HSCs and SECs was more than 85％.The isolated HSCs and SECs could be cultured with 7-10 passages.Conclusion This isolation method is easy and reliable to separate HSCs,SECs and macrophages from the liver of rats with a high purity. Key words: Rat ;  Hepatic non-parenchymal cells ;  Cell isolation ;  Identification