Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (27): 4941-4946.doi: 10.3969/j.issn.2095-4344.2012.27.001

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Isolation, cultivation and osteogenic induction of guinea pig bone marrow mesenchymal stem cells

Shi Xiao-lu1, Duan Li-fu1, Ma Yan2, Bi Xiao-juan2, Liu Li-zhong1   

  1. 1Otorhinolaryngologic Department, 2Medical Research Center, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, Xinjiang Uyghur Autonomous Region, China
  • Received:2012-02-29 Revised:2012-04-16 Online:2012-07-01 Published:2013-11-01
  • Contact: Liu Li-zhong, M.D., Chief physician, Master’s supervisor, Otorhinolaryngologic Department, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, Xinjiang Uyghur Autonomous Region, China liulizhongno.1@163.com
  • About author:Shi Xiao-lu★, Studying for master’s degree, Otorhinolaryngologic Department, First Affiliated Hospital of Xinjiang Medical University, (now Working in Frontier Defence Corps Hospital of Armed Police Force), Urumqi 830054, Xinjiang Uyghur Autonomous Region, China Lauren84103@sina.com

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Abstract:

BACKGROUND: The culture technique and osteogenic induction of bone marrow mesenchymal stem cells would be further investigated based on flexible and easy harvesting of bone marrow mesenchymal stem cells.
OBJECTIVE: To investigate the practical methods of isolation and cultivation of guinea pig bone marrow mesenchymal stromal cells in vitro and to investigate the phenotype characteristics and multilineage differentiation potential of bone marrow mesenchymal stem cells.
METHODS: Guinea pig bone marrow mesenchymal stem cells were isolated and purified by adhesion method. After subculturing and proliferation, expression of cell surface molecule CD29, CD44, CD45 was detected by flow cytometry. The multilineage differentiation capability of bone marrow mesenchymal stem cells was examined by culturing cells under conditions favorable for adipogenic and osteogenic differentiation.
RESULTS AND CONCLUSION: The primary cultured bone marrow mesenchymal stem cells adhered to plastic surface after 96 hours and exhibited an oval, asteroid or fusiform shape. After 8 days, the cells reached 90 % confluence in a single layer. After subculturing and amplification, the cells were further purified with a uniform firbroblast-like morphology. Flow cytometry analysis showed that the positive rate of CD29 and CD44 was 95.7% and 65.7% respectively. After culture in adipogenic and osteogenic medium respectively, passage 3 cells were successfully induced to differentiate into adipogenic and osteogenic lineages, as identified by oil red O, alizarin red S, alkaline phosphatase staining and immunohistochemical type Ⅰ collagenase staining. These findings suggest that by adherence method, in vitro isolated, cultured guinea pig bone marrow mesenchymal stem cells can greatly proliferate and maintain the phenotypic and functional properties of mesenchymal stem cells.

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