Development of reliable primary cultured hepatocytes model / 中国临床药理学与治疗学

ABSTRACT

AIM: To develop a reliable primary cultured hepatocytes model in vitro for liver metastasis research. METHODS: Hepatocytes were isolated by a modification of the two-step collagenase perfusion method. The apoptosis and cell cycle of hepatocytes were measured with flow cytometry. The proliferation of hepatocytes was detected by SRB method. RESULTS: The viability and purity of hepatocytes were 90% and 95%,respectively. The result of flow cytometry analysis showed that there was little apoptosis in hepatocytes and most of hepatocytes were in G_0/G_1 phase. The proliferation and albumin-secreting function of hepatocyte cultured by low glucose DMEM and high glucose DMEM were higher than that of cultured by RPMI1640 during 1 to 6 day, but there was no significant different between low glucose DMEM group and high glucose DMEM group. CONCLUSION: Hepatocytes have higher purity and viability with the normal biological activity for about 6 days by this method and it may be a cell model for the study of liver metastasis in vitro.