Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (6): 1006-1008.doi: 10.3969/j.issn.1673-8225.2009.06.011

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Effect validation of bone marrow-derived mesenchymal stem cells cultured by adherent method in vitro

Zhang Yan, Chen Xi-hai, Ji Yan-chao, Zhai Zhe, Wu Bo   

  1. Department of General Surgery, Fourth Hospital of Harbin Medical University, Harbin  150001, Heilongjiang Province, China
  • Online:2010-02-05 Published:2010-02-05
  • Contact: Wu Bo, Master, Professor, Master’s supervisor, Department of General Surgery, Fourth Hospital of Harbin Medical University, Harbin 150001, Heilongjiang Province, China jbjsbr@126.com
  • About author:Zhang Yan, Master, Department of General Surgery, Fourth Hospital of Harbin Medical University, Harbin 150001, Heilongjiang Province, China
  • Supported by:

    the Tackle Key Program of Heilongjiang Province, No. GC09C409-1*;
    Educational Commission of Heilongjiang Province, No.11541147*

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Abstract:

BACKGROUND: A small number of mesenchymal stem cells (MSCs) present in bone marrow, which would gradually drop with age. 

OBJECTIVE: To verify the effect of adherent method on culture of bone marrow-derived MSCs.

METHODS: Under anaesthesia, bone marrow cells were obtained from femur and tibia of rats, cultured by DMEM containing calf serum, placed in an incubator containing 5% CO2 at 37 ℃. The culture medium was renewed after 24 hours, and remained periodical medium change with once per week. The weakly adherent cells were passaged. The cell morphology, growth curve, and the expression of cell-surface markers were identified by flow cytometry and immunocytochemical staining.

RESULTS AND CONCLUSION: After 24 hours of culture, the cells could adhere to the walls with fusiform or triangle shapes, proliferated faster after 2-3 days, and presented whirlpool-like or clustering. The cells reached a logarithmic growth phase after 2 days, and into the late stationary phase after 12 days, which covered the bottle after 15 days. The cultured cells were positive to CD90 and CD54. The results verified that bone marrow-derived MSCs can be isolated by adherent method. This method is easy operation, and can maintain cell activity preferably.

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