《Chinese Journal of Rehabilitation Theory and Practice》 ›› 2018, Vol. 24 ›› Issue (3): 269-276.doi: 10.3969/j.issn.1006-9771.2018.03.004

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Comparison of Inducing Pluripotent Stem Cells to Differentiate into Motor Neuron Precursor in Two Kinds of System

LI Zhe1, FANG Ming-zhu1, CHEN Hong2, GUO Gang-hua1, FAN Jia-hong1, MAO Zhi-juan2   

  1. 1. Department of Rehabilitation Medicine, The Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, China;
    2. Tongji Hospital, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China
  • Received:2017-11-13 Revised:2018-01-11 Published:2018-03-25 Online:2018-03-27
  • Contact: LI Zhe. E-mail: lizhe.1974@163.com
  • Supported by:
    Supported by National Natural Science Foundation of China (General) (No. 81471302) and Henan Science and Technology Department Basic Research Plan (No. 142300410035)

Abstract: Objective To induce human-induced pluripotent stem cells (iPSCs) to differentiate into spinal motor neuron precursor (MNP) and compare the induction efficiency in systems of feeder and feeder-free. Methods iPSCs cultured on mouse feeder cells or in feeder-free condition were induced into neuroepithelial progenitors (NEP) on the sixth day and MNP on the twelveth day. Their morphology was observed under inverted microscope, and the markers of iPSCs, NEP, MNP were detected with immunofluorescence. NEP-related genes SOX1 and HOXA3, MNP-related genes OLIG2 and PAX6, and pluripotency genes SOX2 and OCT4 were detected with real-time quantitative polymerase chain reaction. Results iPSCs expressed pluripotency markers, while NEP and MNP expressed high levels of neural related markers and low levels of pluripotency markers in two systems. The expression of the genes SOX1, HOXA3, OLIG2 and OCT4 was higher in the feeder system, and there was no significant difference in the expression of genes SOX2 and PAX6.Conclusion iPSCs can differentiate into MNP in culture systems of feeder and feeder-free, and the induction efficiency is higher in the feeder system.

Key words: human-induced pluripotent stem cells, motor neurons, neural differentiation

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