Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (32): 5997-6000.doi: 10.3969/j.issn.1673-8225.2010.32.023

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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

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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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