人胎肝干细胞的体外分离培养与鉴定

Isolation, culture and identification of human fetal liver stem cells In vltro

AIM: To isolate, culture and identify human fetal liver stem cells in vitro to lay a foundation for the application of these cells in bioartificial liver support system and hepatocyte transplantation. METHODS: The fetal liver stem cells were isolated from the liver of dead human fetus. The cell suspension was centrifuged at 50 g. The sediment was resuspended with F12 solution, followed by Percoll gradient centrifugation. The cell layer between 50% and 70% of Percoll was collected, washed, centrifuged, resuspended in DMEM (2 :1), and incubated. Immunohistochemistry was used to identify the phenotypes of the cultured cells. RESULTS: The primary cells were round or oval, much smaller than hepatocytes with larger nucleus and less plasma. These cells grew prosperously in culture medium. The plasma of the cultured cells was positive for CK19, OV6 and albumin staining. Double staining showed that some cells were positive for both albumin and CK19. CONCLUSIONS: The liver stem cells exist in the human fetal liver and display various differentiation status.