A modified method for isolation and purification of human hepatic stellate cells

Objective To improve the method for isolation and purification of human hepatic stellate cells (HSCs). Methods A commonly used method for isolation of human HSCs was modified. The new method only involved type Ⅳ collagenase perfusion of the liver in which hepatic cells were removed by low speed centrifugation and the hepatic nonparenchymal cells were plated and the culture medium was changed twice within 24 h. The cells were subcultured in primary culture in 7-10 d. The morphology and protein marker of the cells were observed at different intervals in the cultures with phase contrast microscopy and immunocytochemical staining. Results The purity of the isolated HSCs was almost 100% in 24 h after first subculture. Conclusion The modified method for the isolation of human HSCs is simple and effective.