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Skeletal muscle regeneration via the chemical induction and expansion of myogenic stem cells in situ or in vitro

Nature Biomedical Engineering volume 5, pages 864–879 (2021)Cite this article

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Abstract

Muscle loss and impairment resulting from traumatic injury can be alleviated by therapies using muscle stem cells. However, collecting sufficient numbers of autologous myogenic stem cells and expanding them efficiently has been challenging. Here we show that myogenic stem cells (predominantly Pax7+ cells)—which were selectively expanded from readily obtainable dermal fibroblasts or skeletal muscle stem cells using a specific cocktail of small molecules and transplanted into muscle injuries in adult, aged or dystrophic mice—led to functional muscle regeneration in the three animal models. We also show that sustained release of the small-molecule cocktail in situ through polymer nanoparticles led to muscle repair by inducing robust activation and expansion of resident satellite cells. Chemically induced stem cell expansion in vitro and in situ may prove to be advantageous for stem cell therapies that aim to regenerate skeletal muscle and other tissues.

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Fig. 1: A small-molecule cocktail induces myogenic cells from dermal cells.
Fig. 2: Characterization of CiMCs.
Fig. 3: Specific upregulation of myogenic gene expression in CiMCs.
Fig. 4: Enriched dermal myogenic cells contribute to chemical-expanded CiMCs.
Fig. 5: The dermal Pax7+ subpopulation is the main contributor to the expanded CiMCs.
Fig. 6: In vivo engraftment of CiMCs promotes muscle regeneration.
Fig. 7: Drug-loaded nanoparticles promote muscle regeneration.
Fig. 8: Drug-loaded nanoparticles enhance muscle repair by promoting in situ satellite cell expansion.

Data availability

The main data supporting the results in this study are available within the paper and its Supplementary Information. Data for the microarray and scRNA-seq have been deposited in the NCBI Gene Expression Omnibus under accession numbers GSE158690 and GSE158691, respectively. All data generated in this study, including source data and the data used to make the figures, are available at Figshare (https://doi.org/10.6084/m9.figshare.13049690.v1).

Code availability

The custom code used is available at GitHub (https://github.com/junrensia/Jun_et_al_Nature_BME_2020).

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Acknowledgements

We thank M. Conboy and V. Rezek for their technical assistance with the in vivo cell transplantation study and K. Heydari for his assistance with FACS. This work was supported in part by grants from UCLA Broad Stem Cell Research Center, the National Institute of Health (EB012240 HL121450 and R56DE029157 to S.L.), a fellowship from the Agency for Science, Technology and Research (to J. Sia), a fellowship from the National Institute of Arthritis and Musculoskeletal and Skin Diseases of the NIH under the Ruth L. Kirschstein National Research Service Award (T32AR059033 (to J. Soto) and the Medical Scientist Training Program at UCLA (NIH T32 GM008042 to L.K.L.). SEM was performed at the California NanoSystems Institute (CNSI) Electron Imaging Center for NanoMachines (EICN) Shared Resource Facility at UCLA. Confocal laser scanning microscopy was performed at the California NanoSystems Institute (CNSI) Advanced Light Microscopy/Spectroscopy Shared Resource Facility at UCLA. FACS was performed at the UCLA Jonsson Comprehensive Cancer Center (JCCC) and Center for AIDS Research Flow Cytometry Core Facility that is supported by National Institutes of Health awards P30 CA016042 and 5P30 AI028697, and by the JCCC, the UCLA AIDS Institute, the David Geffen School of Medicine at UCLA, the UCLA Chancellor’s Office and the UCLA Vice Chancellor’s Office of Research. Single-cell RNA sequencing was conducted at the UCLA Technology Center for Genomics and Bioinformatics.

Author information

Author notes

  1. These authors contributed equally: Jun Fang, Junren Sia.

Authors and Affiliations

  1. Department of Bioengineering, University of California, Los Angeles, Los Angeles, CA, USA

    Jun Fang, Jennifer Soto, Pingping Wang, LeeAnn K. Li, Yuan-Yu Hsueh & Song Li

  2. Department of Medicine, University of California, Los Angeles, Los Angeles, CA, USA

    Jun Fang, Jennifer Soto & Song Li

  3. Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA

    Junren Sia & Raymond Sun

  4. Beijing Key Laboratory of Bioelectromagnetism, Institute of Electrical Engineering, Chinese Academy of Sciences, Beijing, China

    Pingping Wang

  5. David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA

    LeeAnn K. Li

  6. Division of Plastic Surgery, Department of Surgery, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan

    Yuan-Yu Hsueh

  7. Department of Psychiatry and Biobehavioral Sciences, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA

    Kym Francis Faull

  8. Pasarow Mass Spectrometry Laboratory, Jane and Terry Semel Institute for Neuroscience and Human Behavior, University of California, Los Angeles, Los Angeles, CA, USA

    Kym Francis Faull

  9. Department of Integrative Biology and Physiology, University of California, Los Angeles, Los Angeles, CA, USA

    James G. Tidball

  10. Molecular, Cellular & Integrative Physiology Program, University of California, Los Angeles, Los Angeles, CA, USA

    James G. Tidball

  11. Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA

    James G. Tidball

Authors
  1. Jun Fang
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Contributions

J.F., J. Sia, J.G.T. and S.L. designed the project and experiments and wrote the manuscript. J.F., J. Sia and P.W. performed the myogenic induction with small molecules. J.F. performed the immunofluorescence staining, histological analysis, drug delivery system preparation and animal studies. J.F. and J. Soto performed flow cytometry analysis. J. Sia and R.S. performed microarray analysis and gene expression analysis. J.F., J. Sia and J. Soto performed single-cell sequencing and data analysis. J.F. and K.F.F. performed HPLC analysis. J.F., L.K.L. and Y.-Y.H. performed electrophysiological analysis. All of the authors revised the manuscript and added comments.

Corresponding author

Correspondence to Song Li.

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Competing interests

The innovation related to this study has been filed for patent application by J.F., S.L. and J. Sia. (US Serial No. 30435411.2020).

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Supplementary information

Supplementary Information

Supplementary Figs. 1–16 and Table 1, and captions for Supplementary Videos 1–4.

Reporting Summary

Supplementary Video 1

Spontaneous beating after treatment with FR medium for 4 d.

Supplementary Video 2

Spontaneous beating after treatment with FR medium for 8 d.

Supplementary Video 3

Spontaneous beating after treatment with FR medium for 14 d.

Supplementary Video 4

Spontaneous beating after treatment with FR medium for 16 d.

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Fang, J., Sia, J., Soto, J. et al. Skeletal muscle regeneration via the chemical induction and expansion of myogenic stem cells in situ or in vitro. Nat Biomed Eng 5, 864–879 (2021). https://doi.org/10.1038/s41551-021-00696-y

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