| Objective To establish a stable and effective standard procedure for isolate hepatocytes and kupffer cells of pig. Through co-culture kupffer cells with hepatocyte, to understand the influence of kupffer cells on the growth, metabolism, detoxication and physiologic function. In order to provide the data for the high performance and economic and pragmatic bioartificial liver system. Methods The out situ perfused technique with collagenase IV was used to obtain discrete liver cells from the suckling pig liver , and apply densitygradient centrifugation by percoll fluid to isolate kupffer cells respectively , and compare the Diazepam and Acetaminophen cleaning function and existing time. Results We obtained (3. 5±1. 1) × 10~9 liver cells from each suckling pig, and the cell vitality was more than 90%±2%。 The amount of isolated Kupffer cells from each suckling pig is (2.35±0.5) ×10~8, the purity of kupffer cells is 95%±5%. The hepatocyte grew well in twelve to one than eight to one and sixteen to one and hepatocyte alone in morphous and the group of the twelve to one was better in cleaning function of Diazepam and Acetaminophen. When hepatocyte and kupffer cells were cultured together in the proportion twelve to one , the function of hepatocyte was good and could last two weeks.But When we cultured hepatocyte and kupffer cells together in the proportion eight to one or sixteen to one or hepatocyte alone , the hepatocyte could grow 10 days, but the function could only last one week. The hepatocytes and kupffer cells of suckling pig could be cultured together in the proportion twelve to one , and fulfilled the massive culture of liver cells. Conclusion 1. According to the study, we Established a stable and... |
