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中国临床药理学与治疗学 ›› 2005, Vol. 10 ›› Issue (7): 743-746.

• 研究原著 • 上一篇    下一篇

大鼠肝实质细胞原代培养模型的研究及其功能鉴定

周星辉, 王丙力, 谭焕然   

  1. 北京大学医学部基础医学院药理学系, 北京100083
  • 收稿日期:2005-04-05 修回日期:2005-05-18 出版日期:2005-07-26 发布日期:2020-11-10
  • 通讯作者: 谭焕然, 女, 教授, 研究方向:分子药理学。Tel:010-82802004 E-mail: tanlab@sun.bjmu.edu.cn
  • 作者简介:周星辉, 女, 硕士, 研究方向:分子药理学。
  • 基金资助:
    中国高技术研究发展计划(863计划)资助项目(NO108-08-08-03,2002AA214201)

Development of reliable primary cultured hepatocytes model

ZHOU Xing-hui, WANG Bing-li, TAN Huan-ran   

  1. Department of Pharmacology, School of Basic Medical Sciences, Health Center, Peking University, Beijing 100083, China
  • Received:2005-04-05 Revised:2005-05-18 Online:2005-07-26 Published:2020-11-10

摘要: 目的: 为肝细胞功能及相关的研究提供一种客观的、简便的大鼠肝细胞原代培养模型, 并鉴定肝细胞的功能。方法: 在传统的两步灌流法的基础上进行改良分离培养肝实质细胞;流式细胞术测定原代肝细胞的细胞周期和细胞凋亡情况;SRB 法测定不同培养基对原代肝细胞的生长和功能的影响。结果: 分离得到的肝细胞成活率可达90 %以上, 纯度可达95 %以上;流式细胞术结果表明原代肝细胞大多处于G0 /G1 期, 且基本无凋亡;SRB 法和白蛋白分泌量检测结果显示, 低糖DMEM 组原代肝细胞生长和白蛋白分泌功能在短期内(1 ~ 5 d)与高糖DMEM组没有明显差异;RPMI1640 组肝细胞的生长和功能则明显低于前两组(P <0.05)。结论: 体外培养的肝细胞活力和纯度均较高, 体外培养后细胞功能正常, 是一种实用的体外研究肝细胞良好的细胞学模型。

关键词: 大鼠, 肝细胞, 原代培养, 细胞模型, 肿瘤

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 perfusionmethod. 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 G0 /G1 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.

Key words: rats, hepatocyte, primary culture, cell model, tumor

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