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陈钟，丁义涛，南京大学 医学院附属鼓楼医院肝胆外科 江苏省南京市 210008 张鹤云，南京大学生化系 江苏省南京市 210093 陈钟，男，1963-07-04生，江苏省靖江市人，汉族.1983年南通医学院医疗系本科毕业，1988年南通医学院外科硕士研究生毕业，1999年南京大学医学院肝胆外科博士生，副教授，副主任医师，主要从事肝胆外科及生物人工肝研究.江苏省卫生厅重点发展项目基金资助课题 No. BQ200020 项目负责人陈钟，210008，江苏省南京市中山路321号，南京大学医学院附属鼓楼医院肝胆外科. zchen9999@sina.com 电话: 025-3304616 传真：025-6635721 收稿日期 2001-12-04 接受日期 2002-01-04
Serum-free medium for culture of suckling pig hepatocytes
Zhong Chen, Yi-Tao Ding, He-Yun Zhang
Zhong Chen, Yi-Tao Ding, Department of Hepatobiliary Surgery, Affiliated Drum Tower Hospital, Medical School, Nanjing University, Nanjing 210008, Jiangsu Province, China He-Yun Zhang，Department of Biochemistry, Nanjing University, Nanjing 210093, Jiangsu Province, ChinaSupported by the Key Development Foundation of Health Departmentof Jiangsu Province, No. BQ200020 Correspondence to:Zhong Chen, Department of Hepatobiliary Surgery, Affiliated Drum Tower Hospital, Medical School, Nanjing University, Nanjing 210008, Jiangsu Province, China. zchen9999@sina.com Received 2001-12-04 Accepted 2002-01-04
Abstract
AIM:To compare the effect of serum-free medium with serum-contained medium for culture of sucking pig hepatocytes and to search for a kind of better serum-free medium for bioartificial liver.
METHODS:Suckling pig hepatocytes were isolated by modified two-step in situ collagenase perfusion method and cultured at 5×108·L-1 both in serum-free medium and serum-contained medium, respectively. Viability, protein and glucose syntheses, G-6-Pase activity, diazepam transformation of hepatocytes and LDH release were evaluated during 7d of culture.
RESULTS:There were no significant differences in above indices except for lower G-6-Pase activity on 0, 1, 2d and glucose synthesis on 2,7d. High viability (>88%) was obtained though it declined with time. Protein synthesis of hepatocytes cultured in serum-free medium was stable over the course of 7d. Diazepam transformation of hepatocytes was stronger on 2d and 3d (diazepam concentration was 75±4pg·Cell -1 on 2d and 77±5pg·cell-1 on 3d). Glucose synthesis of hepatocytes declined from 1.00±0.15nmol·cell-1 on 1d to 0.71±0.02nmol·cell-1 on 2d. G-6-Pase activity of the hepatocytes declined apparently from 1290±30zmol·cell-1 on 0d to 306±38zmol·cell-1 on 1d, and it was maintained at low level for 7d. LDH release was higher on 2d and 3d.
CONCLUSION:The serum-free medium may be applied in the culture of porcine hepatocytes of bioartificial liver.
Chen Z, Ding YT, Zhang HY. Serum-free medium for culture of suckling pig hepatocytes. Shijie Huaren Xiaohua Zazhi 2002;10(3):320-323
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