Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (6): 968-972.doi: 10.3969/j.issn.1673-8225.2012.06.004

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Isolation, culture and adipogenic ability of different sources of bone marrow derived mesenchymal stem cells

Zhao Zhi-zheng, Lin Hong-sheng   

  1. Department of Oncology, Guang’anmen Hospital, China Academy of Chinese Medical Sciences, Beijing  100053, China
  • Received:2011-12-01 Revised:2011-12-31 Online:2012-02-05 Published:2012-02-05
  • Contact: Lin Hong-sheng, Chief physician, Doctoral supervisor, Department of Oncology, Guang’anmen Hospital, China Academy of Chinese Medical Sciences, Beijing 100053, China drlinhongsheng@163.com
  • About author:Lin Hong-sheng, Chief physician, Doctoral supervisor, Department of Oncology, Guang’anmen Hospital, China Academy of Chinese Medical Sciences, Beijing 100053, China drlinhongsheng@163.com
  • Supported by:

    the National Natural Science Foundation of China, No.30772867*

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Abstract:

BACKGROUND: At present, the most commonly used bone marrow derived mesenchymal stem cells (BMSCs) are murine cells. However, the reports on the purification, expression of surface antigen and the differentiation ability of the BMSCs from rat and mice are rare. 
OBJECTIVE: To isolate, identify and purify different sources of BMSCs and to compare the difference on adipogenic differentiation.
METHODS: The BMSCs were separated from SD rats and Balb/c mice by using differential adhesion, purified by passage, and the purification and proliferation of BMSCs through the process of status were observed. The expression of cell surface antigen of two kinds of BMSCs was identified by flow cytometry. The adipogenic differentiation ability between different sources of BMSCs was compared. 
RESULTS AND CONCLUSION: During isolation and culture of BMSCs, the proliferation of BMSCs from SD rats was significantly stronger than that from Balb/c mice (P < 0.05); the half of the growth period was 36 hours. The typical swirling posted wall growth characteristics could be seen in the third generation. Balb/c mice and SD rats could not express hematopoietic stem cells marker CD34 detected by flow cytometry, and there was no expression of the CD34 cell marker in the subsequent generations (< 5%), with the continuous passage purification, the number of CD29 expression was steady increased, the purity of the surface marker reached to 99% after SD rats BMSCs passaged to the third generation. But the increasing of CD29 was not very obvious in Balb/c mice BMSCs. This fact is consistent with endothelial like cells under the microscope. In adipogenic, SD rats BMSCs still showed a stronger multi-ability; the time for the lipid droplets in SD rats was significantly earlier than that in Balb/c mice. With the passage number increased, the spontaneous adipogenic differentiation phenomenon could be seen in two kinds of primary cells, and the adipogenic differentiation in SD rats BMSCs were stronger than that in the Balb/c mice BMSCs. It was easier to obtain high purity and proliferative source of cells from SD rats BMSCs. The adipogenic capacity of SD rats BMSCs was stronger than that of Balb/c mice BMSCs. 

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