Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (32): 5923-5927.doi: 10.3969/j.issn.1673-8225.2011.32.008

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Culture condition and biological characteristics of rat bone marrow mesenchymal stem cells by using the whole bone marrow adherence method

Zhao Lin1, Feng Zhi-hui1, Jiao Shu-xian1, Li Nan-nan2   

  1. 1Institute of Transfusion Medicine, Qingdao Blood Center, Qingdao  266071, Shandong Province, China
    2Dalian Medical University, Dalian  116027, Liaoning Province, China
  • Received:2011-02-05 Revised:2011-05-16 Online:2011-08-06 Published:2011-08-06
  • Contact: Feng Zhi-hui, Master, Attending physician, Institute of Transfusion Medicine, Qingdao Blood Center, Qingdao 266071, Shandong Province, China fzhheadhunter@163.com
  • About author:Zhao Lin, Chief physician, Institute of Transfusion Medicine, Qingdao Blood Center, Qingdao 266071, Shandong Province, China zhaolinqd@163.com
  • Supported by:

    the Qingdao Science and Technology Support Plan Project, No. 09-1-1-25-nsh*

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Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are widely used as seed cells in tissue engineering and other fields. Therefore, well growth and enough number of rat BMSCs are important for above-mention studies.
OBJECTIVE: To explore the suitable primary culture condition of rat BMSCs in vitro, and to observe the growth property of BMSCs.
METHODS: BMSCs were separated using whole bone marrow culture method and purified using attachment method. Morphology and growth characteristics were observed under inverted phase contrast microscope and cytoskeleton was detected by immunofluorescence. Growth curve was drawn using MTT method. Expression of BMSC surface antigens was detected by flow cytometer.
RESULTS AND CONCLUSION: After isolation and culture, cells developed a spindle or polygonal appearance. Growth curve was S-shaped, and the cultured BMSCs possessed normal morphology and cytoskeleton. At the third passage, BMSCs were positive for CD29, CD44, CD71, CD90, but negative for CD13, CD34, CD45, CD133. Cells isolated and cultured by the adherence separation method in this study are identified as rat BMSCs by morphology and surface antigens, which have the biological characteristics of BMSCs.

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