中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (32): 5997-6000.doi: 10.3969/j.issn.1673-8225.2010.32.023

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

人脂肪间充质干细胞的分离培养及其鉴定

殷莉波1，2，赵文秀2，尹震宇2，杨中萌1，2，张 琼3，王效民2   

  1. 1 厦门大学医学院临床医学系，福建省厦门市 361005；2 厦门大学附属中山医院肝胆外科，福建省厦门市 361004； 3 厦门大学生命科学院，福建省厦门市 361005
  • 出版日期:2010-08-06 发布日期:2010-08-06
  • 通讯作者: 王效民，博士，主任医师，厦门大学附属厦门中山医院肝胆外科 福建厦门市361004 wxm@xmzsh. com
  • 作者简介:殷莉波★，男，1982年生，江苏省江阴市人，汉族，厦门大学医学院外科学在读硕士，主要从事干细胞基础方面的研究。 yinlibo001@163.com
  • 基金资助:

    福建省自然基金（2006J0122）课题名称：肝干细胞的基础研究。国家自然科学基金（30872484）。课题名称：骨髓间充质干细胞向肝脏星状细胞定向分化的构建及其在诱导同种异体胰岛细胞免疫耐受中的作用。

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

Yin Li-bo 1,2, Zhao Wen-xiu2, Yin Zhen-yu2, Yang Zhong-meng 1,2, Zhang Qiong3, Wang Xiao-min2   

  1. 1 Clinical Department, Medical College of Xiamen University, Xiamen  361005, Fujian Province, China; 2 Department of Hepatobiliary Surgery, Zhongshan Hospital, Xiamen University, Xiamen  361004, Fujian Province, China; 3 School of Life  Science, Xiamen University,Xiamen  361005, Fujian Province, China
  • Online:2010-08-06 Published:2010-08-06
  • Contact: Wang Xiao-min, Doctor, Chief physician, Department of Hepatobiliary Surgery, Zhongshan Hospital, Xiamen University, Xiamen 361004, Fujian Province,China wxm@xmzsh.com
  • About author:Yin Li-bo★, Studying for master’s degree, Clinical Department, Medical College of Xiamen University, Xiamen 361005, Fujian Province, China; Department of Hepatobiliary Surgery, Zhongshan Hospital, Xiamen University, Xiamen 361004, Fujian Province, China
  • Supported by:

    the Natural Foundation of Fujian Province, No. 2006J0122; the National Natural Science Foundation of China,  No. 30872484

PDF

707

被引次数

26

摘要：

背景：人脂肪间充质干细胞在不同的条件下被成功诱导分化为成骨细胞、软骨细胞、骨骼肌细胞、肝脏细胞等中胚层细胞，同时也可以向其他胚层分化，证明脂肪间充质干细胞是多分化潜能干细胞。
目的：探讨人脂肪间充质干细胞的分离、鉴定方法。
方法：整形外科吸脂术获得腹部皮下脂肪，0.075%Ⅰ型胶原酶消化，贴壁培养获得人脂肪间充质干细胞，绘制细胞增殖曲线，进行流式细胞、细胞周期、免疫荧光等检测，并验证其是否具有多功能分化潜能。
结果与结论：获得形态较均一的长梭形的人脂肪间充质干细胞，细胞增殖良好，有明显的指数生长期，体外能长期培养存活，并保持不分化状态；流式检测细胞高表达CD105、CD90，低表达CD34、CD45；多数细胞周期检测G0/G1期占80%以上。能向成脂、成骨诱导分化。结果表明，实验分离获得的细胞是具有脂肪间充质干细胞特性的细胞，为深入研究人脂肪间充质干细胞打下基础。

关键词: 脂肪间充质干细胞, 干细胞, 分离, 培养, 鉴定

Abstract:

BACKGROUND: Human adipose tissue-derived mesenchymal stem cells (hADSCs) can differentiate toward mesoblast cell lineage under certain conditions, such as the osteogenic, chondrogenic, myogenic and hepatic lineages, and also can differentiate toward epiblastic and endoblastic layers. Those studies demonstrated that the hADSCs have the capacity of multidifferentiation.
OBJECTIVE: To study isolation and identification of hADSCs.
METHODS: Human adipose tissue was collected after liposuction surgery from the cosmetic and plastic surgery center, digested with 0.075 % collagenase I. Adherent culture could obtain hADSCs. Cell proliferation curve was drawn, and flow cytometry, cell cycle test and immunofluorescence test were conducted to verify whether hADSCs have multiple functional differentiation potential.
RESULTS AND CONCLUSION: hADSCs exhibited fibroblast-like morphology, well proliferation, with obvious exponential growth phase, and could survive in vitro for a long time and maintain a undifferentiated state.  Flow cytometry has shown that cells were highly positive for CD90, CD105, and weakly positive for CD34 and CD45,almost 80% cells stayed in resting state(G0/G1). Cells could differentiate into osteogenic and adipogenic direction. Results have indicated that isolated stem cells show the typical characterization of hADSCs, which is a basis for further studying hADSCs.

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