Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (14): 2471-2475.doi: 10.3969/j.issn.1673-8225.2010.14.001

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Differentiation of rhesus monkey mesenchymal stem cells into neuron-like cells by sonic hedgehog factor

Song Ge 1,2, Zhang Yang3, Liu Bing-qian2, Zheng Wei-wei1, Sun Xue-rong2   

  1. 1 Guangdong Provincial Family Planning Research Institute of Science and Technology, Guangzhou  510600, Guangdong Province, China; 2 State Key Laboratory of Ophthalmology of Sun Yat-sen University, Guangzhou   510060, Guangdong Province, China; 3 the 458  Hospital of Chinese PLA, Guangzhou  510602, Guangdong Province, China
  • Online:2010-04-02 Published:2010-04-02
  • About author:Song Ge☆, Doctor, Associate investigator, Guangdong Provincial Family Planning Research Institute of Science and Technology, Guangzhou 510600, Guangdong Province, China; State Key Laboratory of Ophthalmology of Sun Yat-sen University, Guangzhou 510060, Guangdong Province, China songgepp@126.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30500555*; the Key Program of Family Planning Committee of Guangdong Province, No. 2008001*

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Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) differentiating into neural cells is an effective way of cell therapy of nervous system disease. However, the methods used nowadays still need to be improved.
OBJECTIVE: To induce the differentiation of rhesus monkey BMSCs into neuron-like cells by using sonic hedgehog factor.
METHODS: Rhesus monkey BMSCs differentiating into neuron-like cells was induced by typical retinoic acid and sonic hedgehog factor. Rhesus monkey BMSCs were isolated and cultured by density gradient centrifugation method. Cell growth was observed under an inverted phase contrast microscope and cell growth curve was determined using MTT assay. Flow cytometry was performed to characterize the phenotype of BMSCs, and immunohistochemistry was utilized to assess differentiated cells. Ultra-structure of the differentiated cells was observed by transmission and scanning electron microscopes.
RESULTS AND CONCLUSION: Rhesus monkey BMSCs cultured in vitro were identified by flow cytometry, with high homogenicity. Following sonic hedgehog factor disposal for 7 days, differentiated cells were mainly positive for neurone specific enolase, neurofilament protein, Tau and glial fibrillary acidic protein (GFAP). Image statistical analysis found that in sonic hedgehog factor scheme, neural stem cells marker Nestin positive rate was significantly higher compared with the retinoic acid scheme (P < 0.01). GFAP-positive rate was greater in the retinoic acid scheme than in the sonic hedgehog factor scheme (P < 0.05). Results indicated that sonic hedgehog factor scheme is an effective pathway of rhesus monkey BMSC differentiation into neuron-like cells.

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