Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (6): 973-978.doi: 10.3969/j.issn.1673-8225.2010.06.005

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Human amniotic epithelial cells-secreted neurotrophic factors induces the differentiation of human umbilical cord blood mesenchymal stem cells into neuron-like cells: Possibility verification

Zhang Xiao-ming1, Sun Hai-mei1, Yang Hui2, Ji Feng-qing1   

  1. 1Department of Histology and Embryology,
    2Institute for Neuroscience, Capital Medical University, Beijing   100069, China
  • Online:2010-02-05 Published:2010-02-05
  • Contact: Ji Feng-qing, Doctor, Associate professor, Department of Histology and Embryology, Capital Medical University, Beijing 100069, China jfq@ ccmu. edu.cn
  • About author:Zhang Xiao-ming★, Studying for master's degree, Department of Histology and Embryology, Capital Medical University, Beijing 100069, China zxmailb@sohu.com
  • Supported by:

    the Scientific Research Fund Projects of Beijing Educational Council, No. KM200610025005*;
    the Beijing Municipal Natural Science Foundation, No. 7082017*

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Abstract:

BACKGROUND: Group pre-test has confirmed that amnion endothelial cell conditioned medium can induce human umbilical cord blood mesenchymal stem cells into dopaminergic neuron-like cells. In this process, neurotrophic factors and their receptors may play an important role.

OBJECTIVE: To study the function of neurotrophic factors secreted by amniotic epithelial cells in the differentiation of human umbilical cord blood mesenchymal stem cells into neurons.

METHODS: P1 human umbilical cord blood mesenchymal stem cells at 2×108 /L were incubated and assigned to 3 group. Control group was added with HG-DMEM medium. Induction group received human amniotic epithelial cell medium. Blocking agent group underwent blocking agent K252a fluid, and the incubated was conducted at 36 ℃ for 40 minutes, and then amniotic epithelial cell medium was added. Immunofluorescence chemistry was used to determine neuron specific enolase and dopamine transporter expression in human umbilical cord blood mesenchymal stem cells. Real-time quantitative PCR was employed to detect neuron specific enolase, dopamine transporter and tyrosine hydroxylase expression in human umbilical cord blood mesenchymal stem cells.

RESULTS AND CONCLUSION: Nerve growth factor and brain-derived neurotrophic factor were observed in human amniotic supernatant. P1 human umbilical cord blood mesenchymal stem cells expressed Trka and Trkb. Forty-eight hours following induction, compared with the control group, positive expression of neuron specific enolase and dopamine transporter was significantly increased in the induction and blocking agent groups (P < 0.05), especially in the induction group (P < 0.05). Neuron specific enolase, dopamine transporter and tyrosine hydroxylase mRNA levels were significantly greater in the induction and blocking agent groups compared with the control group (P < 0.01), and each gene mRNA levels were significantly greater in the induction group than in the blocking agent group (P < 0.01). Results verified that neurotrophic factor in the human amniotic epithelial cells plays important effects on differentiation of human umbilical cord blood mesenchymal stem cells into neurons. The promotion effects are mediated by activating Trk receptor.

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