| Objective: To investigate more economical and steady method for the isolating and culturing of hepatic stellate cells (HSCs).Methods: The method for isolating of normal rat HSC was established with collagenase in situ liver recirculation perfusion, and 18%Nycodenz density gradient centrifugation. HSCs were trained in FETAL BOVINE SERUM-DMEM culture medium. Desmin, α-SMA and ICAM-1 were examined for identifing the cells. The immunocytochemical staining was emploied for Collagen and Laminin(LN).Results: The quantity of HSC was above 3.5×10~7 cells per rat. The purity and the viability of the cells were all about 96～98 percent. Besides, the activated HSC expressed LN, collagenⅠand Ⅲ.Conclusion: The result showed that this method of isolating HSC was more economical and steady; the relatively steady cell line was acquired by subculturing. The activated HSC can synthesis more ECM than unactivated HSC.Key Words: HSC,Isolation,Identification... |
PDF Full Text Request