Improvement in isolation and purification of SD rat hepatic stellate cells

Objective To investigate the mechanism of matrix accumulation and hepatic fibrosis and to develop an economic,simple and reliable method for isolation of hepatic stellate cells(HSCs) in rats.Methods HSCs were isolated from liver of rat by perfusion with pronase and collagenase,and were separated from other cells by density gradient centrifugation with Optiprep.The purity of HSCs were cultured in vitro.Results The yield of rat HSCs was 5.0×107/per rat.The cell viability was 98% and the purity of cell was 96%.Conclusion This method for isolating and culturing HSCs is simple,practical,stable and reliable,which settles the foundation of further research of the relationship between HSCs with hepatic fibrosis,particularly the research of cellular level and molecular biology level.