中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (10): 1823-1826.doi: 10.3969/j.issn.1673-8225.2011.10.024

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

人胎盘绒毛膜来源间充质干细胞的生物学特性

张睿婷1，韩之波2，3，王  涛4，孟  磊2，3，刘志鹏1，李  奎4，及月茹3，杨洲鑫3，李扬秋1，韩忠朝1，2，3   

  1. 1暨南大学医学院，广东省广州市 510632
    2中国医学科学院血液学研究所泰达生命科学技术研究中心，天津市 300457
    3中国医学科学院北京协和医学院血液学研究所实验血液学国家重点实验室，天津市 300020
    4广州汉氏联合生物科技有限公司，广东省广州市  511400
  • 收稿日期:2010-11-02 修回日期:2010-12-06 出版日期:2011-03-05 发布日期:2011-03-05
  • 通讯作者: 韩忠朝，博士，博士生导师，暨南大学医学院，广东省广州市 510632；中国医学科学院血液学研究所泰达生命科学技术研究中心，天津市 300457；中国医学科学院北京协和医学院血液学研究所实验血液学国家重点实验室，天津市300020 hanzhongchao@hotmail.com
  • 作者简介:张睿婷★，女，1984年生，湖南省郴州市人，汉族，暨南大学医学院在读硕士，主要从事成体干细胞方面的研究。 zhangruiting66@foxmail.com
  • 基金资助:

    课题受广东省科技厅省部产学研结合示范基地项目(广东省干细胞产业化基地，项目编号：2010B091101004)支持。

Biological characteristics of human chorionic-derived mesenchymal stem cells

Zhang Rui-ting1, Han Zhi-bo2, 3, Wang Tao4, Meng Lei2, 3, Liu Zhi-peng1, Li Kui4, Ji Yue-ru3, Yang Zhou-xin3, Li Yang-qiu1, Han Zhong-chao1, 2, 3   

  1. 1School of Medicine, Jinan University, Guangzhou  510632, Guangdong Province, China
    2TEDA Life Science and Technology Research Center, Institute of Hematology, Chinese Academy of Medical Sciences, Tianjin  300457, China
    3State Key Laboratory of Experimental Hematology, Institute of Hematology, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin  300020, China
    4Guangzhou Health & Biotech Co., Ltd., Guangzhou  510632, Guangdong Province, China
  • Received:2010-11-02 Revised:2010-12-06 Online:2011-03-05 Published:2011-03-05
  • Contact: Han Zhong-chao, Doctor, Doctoral supervisor, School of Medicine, Jinan University, Guangzhou 510632, Guangdong Province, China; TEDA Life Science and Technology Research Center, Institute of Hematology, Chinese Academy of Medical Sciences, Tianjin 300457, China; State Key Laboratory of Experimental Hematology, Institute of Hematology, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, China hanzhongchao@hotmail.com
  • About author:Zhang Rui-ting★, Studying for master’s degree, School of Medicine, Jinan University, Guangzhou 510632, Guangdong Province, China zhangruiting66@foxmail.com
  • Supported by:

    Guangdong Science and Technology Department, the Cooperation Project in Industry, Education and Research of Guangdong Province and Ministry of Education of P.R. China (The Industrialization Base of Stem Cell of Guangdong Province, No. 2010B091101004) *

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摘要：

背景：骨髓是间充质干细胞的主要来源，但骨髓中间充质干细胞数量和质量会随着年龄的增长而逐渐降低。因此，寻找一种新的干细胞来源具有重要意义。
目的：分离研究了一种新的间充质干细胞来源——人绒毛膜来源间充质干细胞，并研究其生物学特性。
方法：将胎盘冲洗干净后，分离出脐带和羊膜组织，将剩余的胎盘组织进行原代和传代培养。通过短串联重复序列分析检测细胞是否来源于绒毛膜，用MTT法检测细胞生长方式，流式细胞仪分析细胞表型及干细胞标记，使用不同的诱导分化培养基检测其多向分化的能力。将该细胞与植物血凝素刺激的外周血单个核细胞共培养，用酶联免疫吸附试验检测上清γ-干扰素的水平。
结果与结论：短串联重复序列分析证明所得到的细胞来源于绒毛膜，这种绒毛膜来源的细胞生长呈典型的成纤维细胞形态。细胞表达常见的间充质干细胞表面标记CD90、CD73、CD105、CD44，不表达CD45，CD11b和CD34。同时，细胞也表达Nestin和Sox-2。在不同的条件培养基培养状态下，细胞可向成骨、成脂方向分化。这些结果证明所得到的细胞为人绒毛膜间充质干细胞。这些绒毛膜间充质干细胞能抑制植物血凝素刺激的人外周血单个核细胞分泌γ-干扰素。可见绒毛膜来源的间充质干细胞具有和传统的骨髓来源间充质干细胞具有相似的生物学特性。

关键词: 人绒毛膜间充质干细胞, 生物学特性, 免疫调节, 诱导分化, 表面标记

Abstract:

BACKGROUND: Bone marrow is major source of mesenchymal stem cells (MSCs), but the quantity and quality of MSCs in bone marrow have been shown to be gradually reduced with the increase of age. Therefore, it’s very important to find a new source of stem cells.
OBJECTIVE: To study a new source of MSCs isolated from human chorionic villous and to study its biological characteristics.
METHODS: Umbilical cord and amniotic membrane were isolated from clean placenta; the remaining placenta tissue underwent primary culture and subculture. Tandem repetitive sequence (TRS) analysis was used to detect whether the MSCs obtained from placenta is chorionic villous original. Cell growth pattern was detected by MTT assay. Flow cytometry was applied to analyze cell phenotype and stem cell markers. The different differential medium was used to detect its multi-directional differentiation ability.
The immunomodulatory properties of these chorionic-derived cells were evaluated by coculturing the cells with peripheral blood mononuclear cells stimulated by phytohemagglutinin (PHA), and the level of interferon-γ (IFN-γ) was detected by enzyme-labeled immunosorbent assay (ELISA).
RESULTS AND CONCLUSION: TRS analysis shows that these chorionic-derived cells are derived from human chorionic. They are adherent cells with typical fibroblast morphology. Flow cytometric analysis reveals that human chorionic-derived cells are consistently positive for MSCs labeled with CD73, CD29, CD44, CD90 and CD105, and negative for the hematopoietic markers CD45, CD11b and CD34. The cells can differentiate into osteoblasts and adipogenesis under different conditioned medium cultured conditions. These results proved that these cells are human chorionic-derived MSCs. The human chorionic-derived MSCs can inhibit peripheral blood mononuclear cell stimulated by PHA to secrete γ-interferon.

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