Long-term culture of hepatic oval cells in rats in vitro

Yin Shou-xin; Zhou Zhen; Ma Mei-xue; Yang Jing-bo; Yang Dong-ye

doi:10.3969/j.issn.2095-4344.2014.29.013

Abstract：BACKGROUND: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 cellactivity. <br> OBJECTIVE:To explore a long-term culture method of hepatic oval cells in vitro. <br> METHODS:Partial y hepatectomized rat model was established by using 2AAF/PH. The regenerated liver was digested by col agenase, 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 cellactivities was determined based on morphology and molecular markers. <br> RESULTS AND CONCLUSION:Both hepatocyte marker ALB and biliary epithelial cellmarker 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 cellactivities for a long time.