Improvement in isolation and culture of SD rat heptic stellate cells

Objective To introduce a high efficient,convenient method for isolation and culture of rat hepatic stellate cells (HSCs). Methods Rat HSCs were isolated by modified in situ perfusion with pronase and collagenase followed by density gradient centrifugation with 12% Optiprep. The purity of HSCs was identified by the expressions of desmin and α-SMA using immunocytochemistry method. Results The harvest rate of hepatic stellate cells was 3×107 cells per liver with the purity of 95% and viability of 98%. Cell proliferation and transdifferentiation remained in good condition after 10 generations of serial subcultivation. Conclusion The rat HSCs are successfully isolated and cultured,suggesting our improvement is feasible.