Hepatic oval cells are recognized as stem/progenitor cells currently, however, long-term culture of hepatic oval cells can inevitably result in the loss of cell activity.OBJECTIVE:
To explore a long-term culture method of hepatic oval cells in vitro.METHODS:
Partially hepatectomized rat model was established by using 2AAF/PH. The regenerated liver was digested by collagenase, and hepatic oval cells were isolated and purified by density gradient centrifugation and identified by immunocytochemistry. Hepatic oval cells were cultured in a medium containing epidermal growth factor and leukaemia inhibitory factor. Then epidermal growth factor and leukaemia inhibitory factor were removed after several months, and its ability of maintaining stem/progenitor cell activities was determined based on morphology and molecular markers.RESULTS AND CONCLUSION:
Both hepatocyte marker ALB and biliary epithelial cell marker CK-19 were found after hepatic oval cells were cultured in a medium containing epidermal growth factor and leukaemia inhibitory factor for 4 months. While in the absence of epidermal growth factor and leukaemia inhibitory factor, the expression of fetal liver marker AFP was decreased quickly. These results indicated that hepatic oval cells could expand in a medium containing epidermal growth factor and leukaemia inhibitory factor and maintain stem/progenitor cell activities for a long time.Subject headings:
hepatocytes; epidermal growth factor; leukemia inhibitory factor; cells, culturedFunding:
the National Natural Science Foundation of China, No. 81072037Funding:
Yin SX, Zhou Z, Ma MX, Yang JB, Yang DY. Long-term culture of hepatic oval cells in rats in vitro. Zhongguo Zuzhi Gongcheng Yanjiu. 2014;18(29):4663-4668.