中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (19): 3601-3605.doi: 10.3969/j.issn.1673-8225.2010.19.039

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

人皮肤源性间充质干细胞体外诱导向淋巴细胞分化的可能性

官立萍，余  杰，黄  冰，罗  挺，黄健发，刘  茜，林丽萍，张  敏，李凯婧，陈系古   

  1. 中山大学中山眼科中心眼科学国家重点实验室，广东省广州市  510060
  • 出版日期:2010-05-07 发布日期:2010-05-07
  • 通讯作者: Huang Bing, Associate investigator, Master’s supervisor, State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center of Sun Yat-sen University, Guangzhou 510060, Guangdong Province, China huangbing2000@hotmail.com
  • 作者简介:官立萍，女，1984年生，河南省驻马店市人，汉族，中山大学在读硕士，主要从事干细胞及免疫系统方面的研究。
  • 基金资助:

    广东省科技计划项目（2008A060202008）；
    国家自然科学基金（30640047）。

In vitro differentiation of human skin-derived mesenchymal stem cells into lymphocytes: Possibility evaluation

Guan Li-ping, Yu Jie, Huang Bing, Luo Ting, Huang Jian-fa, Liu Qian, Lin Li-ping, Zhang Min, Li Kai-jing, Chen Xi-gu   

  1. State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center of Sun Yat-sen University, Guangzhou   510060, Guangdong Province, China
  • Online:2010-05-07 Published:2010-05-07
  • Contact: 黄 冰，副研究员，硕士生导师，中山大学中山眼科中心眼科学国家重点实验室，广东省广州市 510060
  • About author:Guan Li-ping, Studying for master’s degree, State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center of Sun Yat-sen University, Guangzhou 510060, Guangdong Province, China xiaoguan1984@163.com
  • Supported by:

    the Science and Technology Planning Project of Guangdong Province, No. 2008A060202008*;
    the National Natural Science Foundation of China, No. 30640047*

PDF

412

被引次数

5

摘要：

背景：研究发现真皮组织中存在间充质干细胞，存在自体移植的可能性。如能在一定条件下将皮肤来源的间充质干细胞诱导分化为淋巴细胞，则可解决许多免疫系统损伤疾病。

目的：探讨人皮肤源性间充质干细胞经体外诱导向淋巴细胞转分化的可能性。

方法：体外培养人皮肤源性间充质干细胞至第14代，流式细胞仪检测细胞表面标志的表达。诱导人皮肤源性间充质干细胞转分化的条件培养基由人淋巴细胞培养上清液和新鲜人皮肤源性间充质干细胞培养液以7∶3比例组成。倒置显微镜观察诱导后细胞形态学变化。流式细胞仪检测诱导后1~8 d细胞表面淋巴细胞标志的表达。流式细胞仪检测诱导后3，6，9 d人皮肤间充质干细胞自身标记表达的变化。

结果与结论：人皮肤源性间充质干细胞稳定表达自身特异性标志CD73，Vimentin等，不表达造血系统特异性标志CD34，CD45等，且低表达HLA-I，完全不表达HLA-DR。诱导3 d后，细胞体积明显增大，细胞增殖速度明显低于诱导前，细胞内和细胞间存在许多折光性强的囊胞样颗粒。除CD45淋巴细胞标志的表达无明显变化外，诱导后1~ 4 d人皮肤源性间充质干细胞CD3，CD19，CD16，CD4，CD8的表达率均随诱导时间的延长呈线性升高趋势，诱导后5~8 d处于平台期。诱导后3，6，9 d人皮肤源性间充质干细胞自身标记CD73，CD34，Vimentin，HLA-DR的表达无变化，HLA-I的表达率随诱导时间的延长逐渐上升。提示在体外人皮肤源性间充质干细胞可以被诱导向淋巴细胞转分化，具有参与免疫系统损伤修复的潜能。

关键词: 淋巴细胞, 转分化, 人皮肤间充质干细胞, 干细胞

Abstract:

BACKGROUND: Previous research has demonstrated that dermal tissue has mesenchymal stem cells, which have a possibility of autologous transplantation. If the mesenchymal stem cells derived from the skin differentiate into lymphocytes under a certain condition, the immune system disease can be solved generally.
OBJECTIVE: To investigate the possibility of differentiation of human skin-derived mesenchymal stem cells into lymphocytes.
METHODS: Surface marker expression was detected in the 14th passage human skin-derived mesenchymal stem cells using flow cytometry. Transdifferentiation medium of human skin-derived mesenchymal stem cells consisted of human lymphocyte supernatant and fresh human skin-derived mesenchymal stem cells based on the ratio of 7: 3. Inverted microscope was employed to observe morphological changes, and flow cytometry was used to detect surface marker expression in the lymphocytes at 1-8 days after induction. Self-marker expression of human skin-derived mesenchymal stem cells was then detected at 3, 6, and 9 days after induction.
RESULTS AND CONCLUSION: Human skin-derived mesenchymal stem cells stably expressed self-specific marker CD73, Vimentin and so on, but did not express specific markers of hematopoietic system, i.e., CD34, CD45 and so on, lowly expressed HLA-I, but did not express HLA-DR at all. At 3 days after induction, the cell volume significantly increased, cell proliferation rate was significantly lower than before induction, and a lot of cystic-like particles with strong refraction were observed in or between cells. The CD45 lymphocyte expression was not significantly changed, but CD3, CD19, CD16, CD4, and CD8 expression rates of human skin-derived mesenchymal stem cells were linearly increased at 1-4 days after induction and stabilized at 5-8 days after induction. In addition, CD37, CD34, Vimentin, and HLA-DR expressions were not changed at 3, 6, and 9 days after induction, but HLA-I expression rate was gradually increased with the prolongation time of induction. This suggested that human skin-derived mesenchymal stem cells can differentiate into lymphocyte and potentially participate in repairing immune system injury.

中图分类号: