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Highly Efficient Derivation of Skeletal Myotubes from Human Embryonic Stem Cells

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Abstract

Human embryonic stem cells (hESCs) are a promising model for the research of embryonic development and regenerative medicine. Since the first hESC line was established, many researchers have shown that pluripotent hESCs can be directed into many types of functional adult cells in culture. However, most of the reported methods have induced differentiation through the alteration of growth factors in the culture medium. These methods are time consuming; moreover, it is difficult to obtain a pure population of the desired cells because of the low efficiency of induction. In this study, we used a lentiviral-based inducible gene-expression system in hESCs to control the ectopic expression of MyoD, which is an essential transcription factor in skeletal muscle development. The induction of MyoD can efficiently direct the pluripotent hESCs into mesoderm in 24 h. The cells then become proliferated myoblasts and finally form multinucleated myotubes in vitro. The whole procedure took about 10 days, with an induction efficiency of over 90 %. To our knowledge, this is the first time that hESCs have been induced into terminally differentiated cells with only one factor. In the future, these results could be a potential resource for cell therapy for diseases of muscle dysfunction.

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Acknowledgements

This research was supported by grants from the Ministry of Science and Technology of China (2011CB965101), the National Science Fund for Distinguished Young Scholars (31025016), the National Natural Science Foundation of China (30971682, 30901019, 31271586), the Ministry of Science and Technology of China (2011CBA01001, 2012AA020503, 2011AA020108), the Fundamental Research Funds for the Central Universities, the Key Construction Program of the National “985” Project (118000+193411801/006), the Research Fund for the Doctoral Program of Higher Education of China (20110101110098).

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The authors declare no potential conflicts of interest.

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Authors and Affiliations

  1. Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, People’s Republic of China

    Lingjun Rao, Lei Bao, Jijun Chen, Lixiazi He, Pengfei Lu, Jiangtao Ren & Lu Wu

  2. Shanghai East Hospital, Tongji University, Shanghai, 200120, People’s Republic of China

    Wenjie Tang & Youzhen Wei

  3. Stem Cell and Developmental Biology Research Center, College of Animal Science, Zhejiang University, 866 Yuhangtang Road, Nongshenghuan Building E#, Room 301, Hangzhou, 310058, People’s Republic of China

    Lingjun Rao, Lei Bao, Jijun Chen, Haide Chen, Lixiazi He, Pengfei Lu, Jiangtao Ren, Lu Wu, Chun Cui & Lei Xiao

  4. Department of Developmental Biology, Liaoning Medical University, Jinzhou, 121001, People’s Republic of China

    Zhidong Luan

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  1. Lingjun Rao
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  2. Wenjie Tang
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  10. Lu Wu
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  11. Zhidong Luan
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  12. Chun Cui
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  13. Lei Xiao
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Correspondence to Lei Xiao.

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Table S1

Primer sequences for the RT-PCR analyses performed in this study. (XLSX 11 kb)

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Rao, L., Tang, W., Wei, Y. et al. Highly Efficient Derivation of Skeletal Myotubes from Human Embryonic Stem Cells. Stem Cell Rev and Rep 8, 1109–1119 (2012). https://doi.org/10.1007/s12015-012-9413-4

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