中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (32): 5953-5958.doi: 10.3969/j.issn.2095-4344.2012.32.012

• 脂肪干细胞 adipose-derived stem cells • 上一篇    下一篇

人脂肪来源间充质干细胞成骨及成软骨的潜能

朱 寅1，徐卫袁2，王文加3，张兴祥2，严 飞2，沙卫平2，朱现玮1   

  1. 1江苏大学临床医学院，江苏省镇江市 212013；
    2张家港市第一人民医院骨科，江苏省张家港市 215600；
    3 张家港市第一人民医院中心实验室，江苏省张家港市 215600
  • 收稿日期:2012-04-25 修回日期:2012-06-11 出版日期:2012-08-05 发布日期:2012-08-05
  • 通讯作者: 徐卫袁，博士，教授，主任医师，张家港市第一人民医院骨科，江苏省张家港市 215600 xwyl43@163.com
  • 作者简介:朱寅★，男，1987年生，江苏省张家港市人，汉族，江苏大学在读硕士，主要从事脊柱外科、创伤骨科和组织工程方面的研究。 Zhuyin871113@qq.com

Osteogenic and chondrogenic differentiation potential of human adipose-derived mesenchymal stem cells

Zhu Yin1, Xu Wei-yuan2, Wang Wen-jia3, Zhang Xing-xiang2, Yan Fei2, Sha Wei-ping2, Zhu Xian-wei1   

  1. 1Clinical Medical College of Jiangsu University, Zhenjiang 212013, Jiangsu Province, China;
    2Department of Orthopedics, First People’s Hospital of Zhangjiagang, Zhangjiagang 215600, Jiangsu Province, China;
    3Central Laboratory, First People’s Hospital of Zhangjiagang, Zhangjiagang 215600, Jiangsu Province, China
  • Received:2012-04-25 Revised:2012-06-11 Online:2012-08-05 Published:2012-08-05
  • Contact: Xu Wei-yuan, M.D., Professor, Chief physician, Department of Orthopedics, First People’s Hospital of Zhangjiagang, Zhangjiagang 215600, Jiangsu Province, China xwyl43@163.com
  • About author:Zhu Yin★, Studying for master's degree, Clinical Medical College of Jiangsu University, Zhangjiagang 215600, Jiangsu Province, China zhuyin871113@qq.com

PDF

402

被引次数

5

摘要：

背景：人脂肪来源间充质干细胞是种具有较强的体外增殖和多系分化能力的成体干细胞，可以从美容吸脂手术中获得，取材方便，原料来源丰富，在生物治疗应用方面蕴藏着巨大的价值。
目的：体外分离、培养人脂肪来源间充质干细胞，探讨其基本生物学特性及成骨成软骨的潜能。
方法：取美容吸脂获得的脂肪组织，采用Ⅱ型胶原酶消化法分离人脂肪来源间充质干细胞并进行体外培养；观察细胞形态、测定细胞周期、流式细胞仪鉴定细胞表面标志；取第3代细胞，分别加入成骨诱导培养基及成软骨诱导培养基行体外成骨及成软骨诱导。
结果与结论：体外培养人脂肪来源间充质干细胞呈纤维样形态，原代细胞24 h内贴壁，培养5~7 d 后开始形成细胞集落；经细胞周期检测显示G0/G1，S和G2/M所占比例分别为(88±2)%，(12±2)%和0.03%。经流式细胞仪检测CD29和CD105呈阳性表达，CD34和CD45呈阴性表达。RT-PCR检测显示，人脂肪间充质干细胞经成骨诱导分化后细胞中骨桥蛋白mRNA呈阳性表达，经软骨诱导分化后细胞中Ⅱ型胶原mRNA呈阳性表达。结果证实，实验成功体外分离、培养人脂肪来源间充质干细胞，其具有向成骨细胞和软骨细胞分化的潜能。

关键词: 脂肪来源间充质干细胞, 成骨细胞, 软骨细胞, 多向分化, 骨桥蛋白, Ⅱ型胶原, 干细胞

Abstract:

BACKGROUND: Human adipose-derived mesenchymal stem cells are a kind of adult stem cells with the high ability of proliferation and multi-lineage differentiation. They can be acquired from the cosmetic liposuction operation with rich sources of raw material and the selections are very convenient. They have a potential value in the field of bioremediation.
OBJECTIVE: To establish a method of isolating and culturing human adipose-derived mesenchymal stem cells and investigate the proficiency of basic biological characteristics and the potential ability of osteogenic and chondrogenic differentiation.
METHODS: The adipose tissue was taken from the liposuction operation, human adipose-derived mesenchymal stem cells were isolated by type II collagen enzyme digestion and then cultured in vitro. Cell morphology was observed, cell cycle was determined, and cell surface markers were identified with flow cytometry. Passage 3 cells were assigned to two induction groups A, B and two control groups A, B. Then the cells were induced to differentiate into osteoblasts and chondrocytes with different culture media in vitro. The differentiated cells were identified by histochemical staining, immunocytochemical staining and RT-PCR.
RESULTS AND CONCLUSION: After in vitro culture, human adipose-derived mesenchymal stem cells showed the desmoids shape. The primary cells adhered to the wall in 24 hours, and then formed the colony after 5-7 days. The passaged cells adhered to the wall in 4-6 hours and maintained the same shape. Cell cycle studies showed that cells at G0/G1, S and G2/M accounted for (88±2)% (88±2)% and 0.03% respectively. The flow cytometry examination showed that the cells were positive for CD29 and CD105 but they were negative for CD45. RT-PCR showed that after osteogenic induction, human adipose-derived mesenchymal stem cells had positive osteopontin mRNA expression; after chondrogenic induction, cells had positive type II collagen mRNA expression. Human adipose-derived mesenchymal stem cells were successfully isolated and cultured and the cells exhibit the potential to differentiate into osteoblasts and chondrocytes.

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