| Objective: To explore the method of isolating rat hepatic stellate cells(HSC) and hepatocytes(HC) at the same time. Methods: The liver was perfused with collagenase to obtain liver cell suspension. Then liver cell suspension was centrifuged at 500pm to get the gradient, another 500pm then get HC. Lastly HSC were separated from other nonparenchymal cells by density gradient centrifugation . HSCs were identified by the expression of Desmin and α-SMA with the immunohistochemical staining method. Results: The yield rate of HSC was about 1-2×10~7 per rat and that of HC was 1-2×10~8 per rat with this method. The purity of HSC was 95-98% and that of HC was about 90%. More than 95% HSCs were positive by Desmin and α-SMA immunohistochemical staining. Conclusion: The method of isolating rat hepatic stellate cells and hepatocytes at the same time is simple, convenient and reliable, which can supply enough HSCs and HCs for the research. |
PDF Full Text Request