Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (23): 4242-4246.doi: 10.3969/j.issn.1673-8225.2011.23.015

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Isolation, culture and identification of rat adipose-derived stem cells

Qin Yi-wu, Zhao Jin-min, Su Wei, Li Xiao-feng, Liu Wei   

  1. Department of Trauma Orthopedics and Hand Surgery, the First Affiliated Hospital of Guangxi Medical University, Nanning  530021, Guangxi Zhuang Autonomous Region, China
  • Received:2011-01-04 Revised:2011-03-15 Online:2011-06-04 Published:2011-06-04
  • Contact: Zhao Jin-min, Doctor, Professor, Chief physician, Department of Trauma Orthopedics and Hand Surgery, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China zhaojinmin@126.com
  • About author:Qin Yi-wu★, Studying for master's degree, Department of Trauma Orthopedics and Hand Surgery, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China qin.yiwu@163.com
  • Supported by:

    Medical and Health Key Research Program of Guangxi, No. 200636*

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Abstract:

BACKGROUND: Adipose tissue contains abundant adipose-derived stem cells (ADSCs), these cells can be cultured in vitro easily and proliferate rapidly. With the potential of multi-directional differentiation, ADSCs are promising to be seed cells of tissue engineering and have attracted much attention.
OBJECTIVE: To establish a method for the isolation and culture of SD rat ADSCs in vitro, and to investigate some biological characteristics and phenotypes.
METHODS: Collagenase Ⅰwas used to digest and isolate ADSCs from SD rat inguinal fat pads. ADSCs were cultured and passaged in vitro, viewed under inverted microscope and were induced toward osteoblasts and adipocytes. Osteogenic differentiation was confirmed by alkaline phosphatase, alizarin red and von Kossa staining, adipogenic differentiation was confirmed by oil red O staining. Growth curve was drawn and cell surface antigens were identified by flow cytometer.
RESULTS AND CONCLUSION: In vitro cultured ADSCs were spindle-shaped and easily proliferated, well-proportioned after passage. Following many times of passages, ADSCs could still maintain strong proliferative ability. The growth curve was “S” shaped. After osteogenic induction, ADSCs showed a positive reaction for alkaline phosphatase, alizarin red and von Kossa staining, while the adipogenetic induction group showed a positive reaction for oil red O staining. The control group showed negative reaction for every staining. ADSCs were positive for CD29,CD44 and CD105, but negative for CD31 and CD45. ADSCs isolated from SD rat inguinal fat pads can be isolated, cultured, passaged and proliferated easily in vitro, and can be induced into osteoblasts and adipocytes under certain conditions. Furthermore, ADSCs expressed related phenotypes of mesenchymal stem cells.

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