动物造模 z-bio 2022-06-14 449

　　Objective: To explore a stable and economical method for the isolation and culture of hepatic stellate cells in gerbils, and to provide technical support for the in-depth study of the cellular mechanism of hepatic fibrosis in gerbils.

　　Methods: Adult male gerbils were harvested, and the liver cells of gerbils were digested by portal vein perfusion with pronase, collagenase and DNase, and the hepatic stellate cells were separated by Nycodenz density gradient centrifugation. Trypan blue exclusion assay was used to identify cell viability, and α-SMA and Desmin immunocytochemical staining was used to identify cell properties.

　　RESULTS: The yield of hepatic stellate cells was 0.5-1×107/liver. The survival rate of hepatic stellate cells was above 90%. After 3 days of primary culture, the positive cells of α-SMA reached more than 75%, and after subculture, the positive cells of α-SMA and Desmin reached 100%.

　　Conclusion: A stable and reliable method for the isolation and culture of hepatic stellate cells in gerbils has been successfully established, which provides technical support for the research on liver-related diseases and the development of prevention and treatment drugs.