中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (49): 9221-9225.doi: 10.3969/j.issn.2095-4344.2012.49.018

• 干细胞与中医药 stem cells and traditional Chinese medicine • 上一篇    下一篇

虫草多糖体外诱导大鼠骨髓间充质干细胞分化为类肝细胞样细胞

刘江凯1，宋雅芳2，刘友章2，杨谕晨2，张久梅2   

  1. 1河南中医学院第一附属医院，河南省郑州市450008；2广州中医药大学第一附属医院，广东省广州市 510405
  • 收稿日期:2012-03-05 修回日期:2012-07-06 出版日期:2012-12-02 发布日期:2012-12-02
  • 作者简介:刘江凯☆，男，1980年生，2009年广州中医药大学毕业，博士，主治医师，主要从事中医药防治慢性肝病的研究。 ljk1008@yahoo.cn
  • 基金资助:

    广东省科技计划项目(2010B030700068)。

Cordyceps polysaccharide induces differentiation of adult rat mesenchymal stem cells into a
hepatocyte lineage in vitro

Liu Jiang-kai1, Song Ya-fang2, Liu You-zhang2, Yang Yu-chen2, Zhang Jiu-mei2   

  1. 1First Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou 450008, Henan Province, China; 2First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • Received:2012-03-05 Revised:2012-07-06 Online:2012-12-02 Published:2012-12-02
  • About author:Liu Jiang-kai☆, M.D., Attending physician, First Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou 450008, Henan Province, China ljk1008@yahoo.cn
  • Supported by:

    Sceience and Technology Program of Guangdong Province, No. 2010B030700068*

PDF

533

被引次数

9

摘要：

背景：如何促进骨髓间充质干细胞向肝细胞完全意义上的转化，以便更有效改善病变肝组织的结构和功能将成为未来相关研究的重中之重。
目的：探讨虫草多糖在体外诱导大鼠骨髓间充质干细胞分化为类肝细胞样细胞的可行性。
方法：采用贴壁法培养Wistar大鼠骨髓间充质干细胞，实验分3组对骨髓间充质干细胞进行诱导分化，虫草组培养液中加虫草多糖进行诱导，终末质量浓度为0.15 g/L；阳性对照组培养液中加肝细胞生长因子和表皮生长因子联合诱导，质量浓度分别为20 μg/L和10 μg/L；空白对照组仅用含体积分数为10%胎牛血清的DMEM培养液。
结果与结论：分离纯化的骨髓间充质干细胞经流式细胞仪检测显示CD34阴性表达，CD44阳性表达。诱导分化7 d时虫草组及阳性对照组细胞出现甲胎蛋白表达，14 d时表达增强，28 d时表达减弱，阳性对照组甲胎蛋白阳性率高于虫草组；诱导分化7 d时阳性对照组细胞角蛋白18出现表达，14 d时表达增强，虫草组则在14 d时出现表达，而后持续。诱导分化7 d时虫草组及阳性对照组白蛋白表达阴性，14 d时出现阳性。诱导分化14 d时虫草组及阳性对照组细胞糖原染色阳性，28 d时表达减弱。空白对照组结果皆为阴性。结果可见虫草多糖可以在体外诱导大鼠骨髓间充质干细胞分化为类肝细胞样细胞。

关键词: 虫草多糖, 骨髓间充质干细胞, 分化, 肝细胞, 甲胎蛋白, 白蛋白, 角蛋白18, 干细胞

Abstract:

BACKGROUND: How to promote the transformation of bone marrow mesenchymal stem cells into hepatocytes to effectively improve the structure and function of diseased hepatic tissue will become a key part of future studies.
OBJECTIVE: To investigate the possibility of cordyceps polysaccharide inducing the differentiation of rat bone marrow mesenchymal stem cells into hepatocyte-like cells in vitro.
METHODS: Wistar rat bone marrow mesenchymal stem cells were cultured by adherence method and then randomized to three groups. In the cordyceps polysaccharide group, bone marrow mesenchymal stem cells were induced with 0.15 g/L cordyceps polysaccharide. In the positive control group, hepatocyte growth factor (20 μg/L) and epidermal growth factor (10 μg/L) were added. In the blank control group, DMEM containing 10% fetal bovine serum was added.
RESULTS AND CONCLUSION: Flow cytometry showed that the isolated and purified bone marrow mesenchymal stem cells were negative for CD34 but positive for CD44. In the cordyceps polysaccharide and positive control group, α-fetoprotein expression appeared at 7 days, peaked at 14 days and was attenuated at 28 days. α-fetoprotein expression was higher in the positive control group than that in the cordyceps polysaccharide group at each time point. In the positive control group, cytokeratin 18 expression appeared at 7 days and was enhanced at 14 days. In the cordyceps polysaccharide group, cytokeratin 18 expression appeared at 14 days and continued thereafter. In the cordyceps polysaccharide and positive control groups, albumin expression was negative at 7 days, but it turned positive at 14 days. In the cordyceps polysaccharide and positive control groups, glycogen staining appeared positive at 14 days but it was attenuated at 28 days, and there was no positive glycogen staining throughout the experimental period   in the blank control group. These findings suggest that cordyceps polysaccharide can induce rat bone marrow mesenchymal stem cells into hepatocyte-like cells. 

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