Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (10): 1884-1888.doi: 10.3969/j.issn.1673-8225.2012.10.038

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Tanshinone-ⅡA induced differentiation of rat bone marrow mesenchymal stem cells into neuron-like cells in vitro after pre-induced by basic fibroblast growth factors

Li Yu-mei1, Yu De-li2, Liu Lai-bing3, Yu Zi-jiang1   

  1. 1Department of Anatomy, Guiyang Medical University, Guiyang  550004, Guizhou Province, China; 2Second Clinical Department, Guiyang University of Traditional Chinese Medicine, Guiyang  550001, Guizhou Province, China; 3Neurosurgery Department, Guiyang University of Traditional Chinese Medicine, Guiyang  550004, Guizhou Province, China
  • Received:2011-06-28 Revised:2011-09-14 Online:2012-03-04 Published:2012-03-04
  • Contact: author: Yu Zi-jiang, M.D., Professor, Department of Anatomy, Guiyang Medical University, Guiyang 550004, Guizhou Province, China yuzijiang@yahoo.cn
  • About author:Li Yu-mei, Teaching assistant, Department of Anatomy, Guiyang Medical University, Guiyang 550004, Guizhou Province, China 136721361@qq.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30660189*, 81060108*; Social Development Foundation of Science and Technology Department of Guizhou Province, No. SY[2009]3058*

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Abstract:

BACKGROUND: Whether bone marrow mesenchymal stem cells (BMSCs) induced by traditional Chinese medicine could differentiate into neuron-like cells remains poorly understood.                             
OBJECTIVE: To identify the efficiency of tanshinone-ⅡA containing traditional Chinese medicine on the differentiation of rat BMSCs into neuron-like cells in vitro after pre-induced by basic fibroblast growth factors.                                            
METHODS: One month old healthy male Wistar rats were used in this experiment, and BMSCs were separated and purified by Percoll medium density gradient centrifugation, repeated subculture and differential adherence methods. Passage 3 BMSCs were differentiated into neuron-like cells induced by basic fibroblast growth factors (bFGF) and tanshinone-ⅡA. All-trans retinoic acid serum-free medium induced cells and non-induced cells served as controls.                                            
RESULTS AND CONCLUSION: ①BMSCs could stably express the stem cells specific markers CD44, and the positive rate was (91.00±1.58)%. And the negative expressions of CD34 and CD45 were observed in the BMSCs by immunohistochemistry and flow cytomtry and the purity of stem cells was up to 95.5%. ② After directional differentiation and cultivation, typical neuron-like cells could be seen under scanning electron microscope. Positive expression of NSE was detected by immunohistochemistry and the possitive rate was (75.6±2.31)%. Nestin expression was gradually decreased to negative as well as the expression of glial fibrillary acidic protein. The results suggested that tanshinone-ⅡA can directly and efficiently induce BMSCs differentiating into neuron-like cells after pre-induced by basic fibroblast growth factors.
 

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