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A medium both for cell growth and avoiding adding serum medium is imminent, so serum-free medium emerge as the times require.To develop a new serum-free medium for C3A cells to lay the foundation for the clinical application of hepatocytes and bioartificial liver.C3A cells were cultivated with serum-free medium, complete medium and basal medium, respectively. The growth and morphological characteristics of cells were observed. Curves of cell growth and the energy were drawn. The leakage volume of alaninetransaminase, urea level and the level of albumin were determined by automatic biochemistry analyzer and radioimmunoassay. The changes of gene expression were determined by real-time fluorescence quantitative PCR.The cells in the three kinds of mediums grew well. Apart from the 7th day, the cell density had no significant difference between the serum-free medium and complete medium groups in the culture period (P > 0.05), whereas it was obviously higher in these two groups than in the basal medium group at 4-7 days (P > 0.05). The level of urea in the serum-free medium group was significantly higher than that in the other two groups, and the level of urea in the complete group was higher than that in the basal medium group (P < 0.05). Real-time fluorescence quantitative RCR revealed that gene expressions of CK18, CK19 and hepatocyte nuclear factor 4α were lower in the serum-free group as compared with the complete medium group (P < 0.05). The results showed that the new serum-free medium for hepatocytes is effective just like the conventional complete medium.Zhao YC, Wang Y, Lu H, Jian GD, Pan MX, Gao Y. A new serum-free medium for hepatocyte growth. Zhongguo Zuzhi Gongcheng Yanjiu. 2012;16(11): 1963-1968.