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Abstract

Muscle regeneration is a dynamic process during which cell state and identity change over time. A major roadblock has been a lack of tools to resolve a myogenic progression in vivo. Here we capitalize on a transformative technology, single-cell mass cytometry (CyTOF), to identify in vivo skeletal muscle stem cell and previously unrecognized progenitor populations that precede differentiation. We discovered two cell surface markers, CD9 and CD104, whose combined expression enabled in vivo identification and prospective isolation of stem and progenitor cells. Data analysis using the X-shift algorithm paired with single-cell force-directed layout visualization defined a molecular signature of the activated stem cell state (CD44+/CD98+/MyoD+) and delineated a myogenic trajectory during recovery from acute muscle injury. Our studies uncover the dynamics of skeletal muscle regeneration in vivo and pave the way for the elucidation of the regulatory networks that underlie cell-state transitions in muscle diseases and ageing.

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Change history

  • 05 March 2018

    In the version of this Article originally published, the name of author Andrew Tri Van Ho was coded wrongly, resulting in it being incorrect when exported to citation databases. This has been corrected, though no visible changes will be apparent.

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Acknowledgements

We thank D. Burns and F. Gherardini for valuable discussion; G. Han for help with graphics; M. Kyba for Pax7-ZsGreen transgenic mice and M. A. Rudnicki for Pax7 knockout mice; K. Koleckar, P. Kraft and M. Blake for technical assistance; and the Stanford Shared FACS Facility for technical support. This study was supported by a BD Biosciences Stem Cell grant (E.P.); US National Institutes of Health (NIH) grant K99AG042491 (B.D.C.); Muscular Dystrophy Association (MDA) development grant 217821 (A.T.V.H.), NIH grants NS089533 and AG020961, California Institute for Regenerative Medicine grant RB5-07469 and the Baxter Foundation (H.M.B.).

Author information

Author notes

    • Benjamin D. Cosgrove
    • , Kara L. Davis
    •  & Sean C. Bendall

    Present addresses: Meinig School of Biomedical Engineering, Cornell University, Ithaca, New York 14853, USA (B.D.C.); Bass Center for Childhood Cancer and Blood Disorders, Lucile Packard Children’s Hospital, Stanford University School of Medicine, Stanford, California 94305, USA (K.L.D.); Department of Pathology, Stanford University, Stanford, California 94305, USA (S.C.B.).

Affiliations

  1. Blau Laboratory, Stanford University School of Medicine, Stanford, California 94305, USA

    • Ermelinda Porpiglia
    • , Andrew Tri Van Ho
    • , Benjamin D. Cosgrove
    • , Thach Mai
    •  & Helen M. Blau
  2. Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, California 94305, USA

    • Ermelinda Porpiglia
    • , Nikolay Samusik
    • , Andrew Tri Van Ho
    • , Benjamin D. Cosgrove
    • , Thach Mai
    • , Kara L. Davis
    • , Astraea Jager
    • , Garry P. Nolan
    • , Sean C. Bendall
    • , Wendy J. Fantl
    •  & Helen M. Blau
  3. Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California 94305, USA

    • Ermelinda Porpiglia
    • , Andrew Tri Van Ho
    • , Benjamin D. Cosgrove
    • , Thach Mai
    •  & Helen M. Blau
  4. Nolan Laboratory, Stanford University School of Medicine, Stanford, California 94305, USA

    • Nikolay Samusik
    • , Kara L. Davis
    • , Astraea Jager
    • , Garry P. Nolan
    •  & Sean C. Bendall
  5. Stanford Comprehensive Cancer Institute and Department of Obstetrics and Gynecology, Stanford University School of Medicine, Stanford California, California 94305, USA

    • Wendy J. Fantl

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Contributions

E.P. and H.M.B. conceived the study. E.P. designed and performed experiments, analysed and interpreted data and wrote the manuscript. N.S. developed the analysis algorithm, and analysed and interpreted data. H.M.B., W.J.F. and A.T.V.H. designed experiments, analysed and interpreted data and wrote the manuscript. T.M., K.L.D., S.C.B., B.D.C. and G.P.N. analysed and interpreted data. A.J. provided technical support with antibody conjugation and CyTOF data acquisition.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Helen M. Blau.

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DOI

https://doi.org/10.1038/ncb3507

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