中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (27): 5067-5070.doi: 10.3969/j.issn.1673-8225.2010.27.029

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

脂肪间充质干细胞的分离培养和鉴定

姚素艳1，杨依勇2，秦书俭2，郑德宇2   

  1. 辽宁医学院，1病理生理学教研室，2解剖学教研室，辽宁省锦州市  121001
  • 出版日期:2010-07-02 发布日期:2010-07-02
  • 通讯作者: 郑德宇，副教授，辽宁医学院解剖学教研室，辽宁省锦州市 121001 zheng_deyu2000@yahoo.com.cn
  • 作者简介:姚素艳★，女，1971年生，辽宁省兴城市人，满族，2005年辽宁医学院毕业，硕士，副教授，主要从事神经退行性变和骨组织工程方面的研究。 ysyzdy@yahoo. com.cn
  • 基金资助:

    辽宁省教育厅创新团队项目(2006T063)。

Isolation, culture and identification of adipose tissue-derived mesenchymal stem cells

Yao Su-yan1, Yang Yi-yong2, Qin Shu-jian2, Zheng De-yu2   

  1. 1 Department of Pathophysiology, 2 Department of Anatomy, Liaoning Medical University, Jinzhou  121001, Liaoning Province, China
  • Online:2010-07-02 Published:2010-07-02
  • Contact: Zheng De-yu, Associate professor, Department of Anatomy, Liaoning Medical University, Jinzhou 121001, Liaoning Province, China zheng_deyu2000@yahoo.com.cn
  • About author:Yao Su-yan★, Master, Associate professor, Department of Pathophysiology, Liaoning Medical University, Jinzhou 121001, Liaoning Province, China ysyzdy@yahoo.com.cn
  • Supported by:

    the Innovation Team Program of Department of Education of Liaoning Province, No. 2006T063*

PDF

796

被引次数

12

摘要：

背景：近年来的研究发现，在脂肪组织中也存在这与骨髓间充质干细胞类似的间充质干细胞。脂肪组织可以从美容吸脂中获得，来源丰富，取材方便，蕴藏着巨大的临床应用价值。
目的：分离培养人脂肪间充质干细胞并检测其生物学特性。
方法：用离心的方法获得人脂肪间充质干细胞，接种于含体积分数为10%胎牛血清的DMEM培养基中行贴壁培养。并进行细胞形态学观察，绘制细胞生长曲线，流式细胞仪检测细胞表面抗原。
结果与结论：采用消化、离心分离获得的脂肪间充质干细胞大小较为均匀，呈梭形或星形的成纤维细胞样。细胞生长曲线测定表明接后第5天细胞进入指数增生期，至第9天后数量减少；流式细胞仪检测表明超过90%细胞为CD29、CD45和CD105阳性。提示体外分离培养脂肪间充质干细胞生长稳定，可作为组织工程的种子细胞。

关键词: 脂肪间充质干细胞, 流式细胞仪, 免疫细胞化学, 分离培养, 鉴定

Abstract:

BACKGROUND: Previous studies have demonstrated that there are similar mesenchymal stem cells in adipose tissue as bone marrow mesenchymal stem cells (BMSCs). Adipose tissue can be obtained from lipid absorption of cosmetology, and is characterized by abundant source and convenient collection, which shows great value in clinical application.
OBJECTIVE: To isolate and culture human adipose tissue-derived mesenchymal stem cells (ADMSCs) and to measure its biological properties.
METHODS: Human ADMSCs were isolated by centrifugation and cultured in DMEM medium containing 10% fetal bovine serum. The morphology of cells was observed, and cell growth curve was drawn. Cell surface antigens were measured using flow cytometry.
RESULTS AND CONCLUSION: ADMSCs obtained by digestion and centrifugation were similar in size, spindle-shaped or star-shaped fibroblasts-liked cells. Cell growth curve analysis indicated that ADMSCs were in the exponential stage at 5 days following incubation. The number of ADMSCs was reduced at 9 days. Flow cytometry analysis showed that more than 90% cells were positive for CD29, CD45 and CD105. These indicated that human ADMSCs isolated in vitro grew stably and can be used as tissue engineered seed cells.

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