中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (36): 6669-6672.doi: 10.3969/j.issn.1673-8225.2011.36.005

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

大鼠骨髓间充质干细胞体外诱导为肝细胞样细胞

张  涛，赵振国，高  辉   

  1. 解放军成都军区总医院肿瘤诊治中心，四川省成都市  610083
  • 收稿日期:2011-01-12 修回日期:2011-04-29 出版日期:2011-09-03 发布日期:2011-09-03
  • 作者简介:张涛★，1963年生，四川省内江市人，汉族，1990年解放军第三军医大学毕业，硕士，主任医师，主要从事肿瘤内科、肝癌干细胞研究工作。 zhangtao269@163.com
  • 基金资助:

    四川省卫生厅课题(070388)，课题名称：骨髓间充质干细胞在二乙基亚硝胺诱导大鼠原发性肝癌模型中的转化研究；四川省科技厅课题(2009JY0016)，课题名称：间充质干细胞在原发性肝癌微环境下的转归研究；成都军区资助课题(MB07013)，课题名称：MSC在DENA诱导大鼠原发性肝癌模型中的转归研究。

Induced differentiation of rat bone marrow mesenchymal stem cells into hepatocyte-like cells in vitro

Zhang Tao, Zhao Zhen-guo, Gao Hui   

  1. Center of Tumor Diagnosis and Treatment, PLA General Hospital of Chengdu Military Region, Chengdu  610083, Sichuan Province, China
  • Received:2011-01-12 Revised:2011-04-29 Online:2011-09-03 Published:2011-09-03
  • About author:Zhang Tao★, Master, Chief physician, Center of Tumor Diagnosis and Treatment, PLA General Hospital of Chengdu Military Region, Chengdu 610083, Sichuan Province, China zhangtao269@163.com
  • Supported by:

    the Health Bureau of Sichuan Province, No. 070388*; the Science and Technology Bureau of Sichuan Province, No. 2009JY0016*; the Funding Project of Chengdu Military Region, No. MB07013*

PDF

334

被引次数

7

摘要：

背景：目前体外诱导骨髓间充质干细胞分化为肝细胞样细胞的研究主要集中在不同诱导因子的诱导作用，而微环境的诱导作用研究较少。
目的：观察肝细胞生长因子、胎肝细胞对骨髓间充质干细胞体外诱导分化为肝细胞样细胞的影响。
方法：采用密度梯度离心法分离大鼠骨髓间充质干细胞，反复贴壁法纯化扩增骨髓间充质干细胞；采用Ⅰ型胶原酶消化法分离孕3周大鼠胚胎肝脏细胞，差速贴壁法纯化肝脏细胞；阴性对照组骨髓间充质干细胞只加入含体积分数为10%胎牛血清的L-DMEM培养基。诱导组在阴性对照组基础上加入一定浓度肝细胞生长因子或者与胎肝细胞共培养进行诱导分化。
结果与结论：诱导组白蛋白、甲胎蛋白水平均高于非诱导组(P < 0.01),诱导组糖原染色阳性，免疫细胞化学染色CK-18阳性，而非诱导组糖原染色、CK-18均阴性。结果显示肝细胞生长因子和胎肝细胞均可在体外诱导大鼠骨髓间充质干细胞分化为具有肝细胞样细胞表型和功能的类肝细胞。

关键词: 肝细胞样细胞, 骨髓间充质干细胞, 胎肝细胞, 肝干细胞, 肝细胞生长因子

Abstract:

BACKGROUND: The study of the induction of differentiation of rat bone marrow mesenchymal stem cells (BMSCs) into hepatocyte-like cells majorly concentrates on the induction of different inducing factors, and pays little attention on the induction of micro environment.
OBJECTIVE: To investigate the differentiation of rat BMSCs into hepatocyte-like cells.
METHODS: The rats BMSCs were isolated by density gradient centrifugation and purified by adherence cultivation, morphology feature of BMSCs were observed by microscope. The fetal liver cells were isolated by adherence cultivation after digesting the rats embryo liver of 3 weeks with collagenase. The negative control group was cultured in L-DMEM containing 10% fetal bovine serum. The purified BMSCs in the induced group were cultured in L-DMEM containing 10% fetal bovine serum with hepatocyte growth factor (HGF) and fetal liver cells.
RESULTS AND CONCLUSION: Compared with those in non-induced BMSCs, the levels of alpha-fetoprotein and albumin in the induced-BMSCs were higher (P < 0.01). Both glycogen and CK-18 were positive in the induced BMSCs. BMSCs can differentiate into hepatocyte-like cells with hepatic phenotype and function in the presence of HGF and fetal liver cells, which may be used as a kind of cell resources to treat severe hepatic disease.

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