中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (49): 9257-.doi: 10.3969/j.issn.1673-8225.2010.49.031

• 干细胞基础实验 • 上一篇    下一篇

体外诱导人脐带间充质干细胞向肝样细胞的分化

陈军1，刘玉侠1，姜文华2，韩光3，姜文平4   

  1. 1吉林省肿瘤防治研究所，吉林省长春市 130012；  2吉林大学基础白求恩医学院组织胚胎教研室，吉林省长春市 130021；3吉林省肿瘤医院医教科，吉林省长春市 130012；4吉林油田总医院核医学科，吉林省松原市138000
  • 出版日期:2010-12-03 发布日期:2010-12-03
  • 通讯作者: 姜文平，吉林油田总医院核医学科，吉林省松原市 138000 jiangwenhua468@163.com
  • 作者简介:陈军，男，1956年生，吉林省长春市人，汉族，1982年长春中医学院毕业，教授，博士生导师，主要从事肿瘤免疫方面的研究。 yuhongnew@yahoo.com.cn
  • 基金资助:

    吉林省科技发展计划项目(200905200)，课题名称“人脐带间充质干细胞诱导分化为肝前体细胞的实验研究”。

Differentiation of human umbilical cord mesenchymal stem cells into hepatocyte-like cells in vitro

Chen Jun1, Liu Yu-xia1, Jiang Wen-hua2, Han Guang3, Jiang Wen-ping4   

  1. 1Jilin Province Institute of Cancer Research, Changchun  130012, Jilin Province, China; 2Department of Histology and Embryology, Bethune School of Basic Medical Sciences, Jilin University, Changchun  130021, Jilin Province, China; 3Department of Medical Teaching, Tumor Hospital of Jilin Province, Changchun  130012, Jilin Province, China; 4Department of Nuclear Medicine, Jilin Oil Field Hospital, Songyuan 138000, Jilin Province, China
  • Online:2010-12-03 Published:2010-12-03
  • Contact: Jiang Wen-ping, Department of Nuclear Medicine, Jilin Oil Field Hospital, Songyuan 138000, Jilin Province, China jiangwenhua468@ 163.com
  • About author:Chen Jun, Professor, Doctoral supervisor, Jilin Province Institute of Cancer Research, Changchun 130012, Jilin Province, China yuhongnew@yahoo.com.cn
  • Supported by:

    the Scientific and Technological Development Project of Jilin Province, No. 200905200*

PDF

353

被引次数

27

摘要：

背景：多项研究表明间充质干细胞具有向肝细胞分化的潜能，这将为肝细胞移植、生物人工肝、肝组织工程提供大量种子细胞，也有望在急性肝衰竭、终末期肝病和遗传代谢性肝脏疾病治疗方面带来较大突破。
目的：拟在体外诱导人脐带间充质干细胞向肝样细胞分化，观察其形态及功能变化。
方法：取体外培养第3代处于对数生长期的人脐带间充质干细胞，以1×109 L-1的细胞浓度种植在培养瓶中，加入含体积分数为10%胎牛血清的DMEM/F12培养基，设立3组：实验1组加入20 μg/L肝细胞生长因子+10 μg/L碱性成纤维细胞生长因子，实验2组在其基础上另加入20 μg/L制瘤素，空白对照组不加入任何生长因子。根据细胞生长情况，每周换液两三次。分别于培养7，14，18 d在倒置显微镜下观察细胞形态，采用免疫细胞化学检测甲胎蛋白、白蛋白及细胞角蛋白18的表达，运用PAS法检测糖原的表达。
结果与结论：人脐带间充质干细胞可在含20 μg/L肝细胞生长因子、10 μg/L碱性成纤维细胞生长因子、20 μg/L制瘤素、体积分数为10%胎牛血清的DMEM/F12培养体系中诱导分化为具有肝细胞表型和功能的细胞，且肝细胞生长因子、碱性成纤维细胞生长因子、制瘤素联合应用的诱导效果优于单纯应用前两者。

关键词: 诱导分化, 肝样细胞, 肝细胞生长因子, 碱性成纤维细胞生长因子, 制瘤素, 间充质干细胞, 脐带, 干细胞培养与分化

Abstract:

BACKGROUND: Many studies have shown that mesenchymal stem cells have the potential to differentiate into hepatocytes, which will not only provide seed cell source for hepatocellular transplantation, bioartificial liver and hepatic tissue engineering, but also open breakthrough therapies in acute liver failure, end-stage liver disease and genetic metabolic liver disease.
OBJECTIVE: To induce human umbilical cord mesenchymal stem cells into hepatocyte-like cells in vitro, to observe morphological and functional changes respectively.
METHODS: The logarithmic phase of human umbilical cord mesenchymal stem cells of P3 was cultured at the density of 1×109/L in culture flasks containing DMEM/F12 with 10% fetal bovine serum (FBS). The cells were divided into 3 groups: group 1 treated with DMEM/F12 medium, containing 10% FBS, 20 μg/L hepatocyte growth factor (HGF), 10 μg/L basic fibroblast growth factor (bFGF), group 2 treated with DMEM/F12 medium, containing 10% FBS, 20 μg/L HGF, 10 μg/L bFGF, 20 μg/L oncostatin, blank control group treated without growth factor. The medium was changed 2-3 times a week according to cell growth. The cell morphology was observed at 7, 14, 18 days following culture under an inverted microscope. Expressions of alpha fetoprotein, albumin and cytokeratin18 were examined by immunocytochemistry. The glycogen deposits were examined by periodic acid-Schiff method.
RESULTS AND CONCLUSION: The human umbilical cord mesenchymal stem cells could be differentiated into heaptocyte-like cells by DMEM/F12 medium supplemented with 10% FBS, 20 μg/L HGF, 10 μg/L bFGF and 20 μg/L oncostatin. The induced efficiency was more than by DMEM/F12 medium supplemented with 10% FBS, 20 μg/L HGF, 10 μg/L bFGF.

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