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Self-renewal and expansion of single transplanted muscle stem cells

Abstract

Adult muscle satellite cells have a principal role in postnatal skeletal muscle growth and regeneration1. Satellite cells reside as quiescent cells underneath the basal lamina that surrounds muscle fibres2 and respond to damage by giving rise to transient amplifying cells (progenitors) and myoblasts that fuse with myofibres. Recent experiments showed that, in contrast to cultured myoblasts, satellite cells freshly isolated3,4,5 or satellite cells derived from the transplantation of one intact myofibre6 contribute robustly to muscle repair. However, because satellite cells are known to be heterogeneous4,6,7, clonal analysis is required to demonstrate stem cell function. Here we show that when a single luciferase-expressing muscle stem cell is transplanted into the muscle of mice it is capable of extensive proliferation, contributes to muscle fibres, and Pax7+luciferase+ mononucleated cells can be readily re-isolated, providing evidence of muscle stem cell self-renewal. In addition, we show using in vivo bioluminescence imaging that the dynamics of muscle stem cell behaviour during muscle repair can be followed in a manner not possible using traditional retrospective histological analyses. By imaging luciferase activity, real-time quantitative and kinetic analyses show that donor-derived muscle stem cells proliferate and engraft rapidly after injection until homeostasis is reached. On injury, donor-derived mononucleated cells generate massive waves of cell proliferation. Together, these results show that the progeny of a single luciferase-expressing muscle stem cell can both self-renew and differentiate after transplantation in mice, providing new evidence at the clonal level that self-renewal is an autonomous property of a single adult muscle stem cell.

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Figure 1: Characterization of the integrin-α 7 + CD34 + fraction as muscle stem cells.
Figure 2: MuSC engraftment monitored by in vivo non-invasive bioluminescence imaging.
Figure 3: MuSC proliferation in response to muscle tissue damage.
Figure 4: Transplantation of a single MuSC demonstrates self-renewal capacity.

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Acknowledgements

We thank O. Alkan for developing the single cell RT–PCR protocol, K. Koleckar, M. Pajcini and T. Doyle for technical support, T. Brazelton, F. Rossi and S. Corbel for comments on the manuscript, J. Ramunas for statistical analysis, and M. Lutolf and P. Gilbert for fabricating the hydrogel microwells. We also thank F. Rossi for providing the integrin-α7–PE-conjugated antibody, and C. Contag for providing the FLuc mice. This work was supported by NIH grants AG009521, AG024987 and by the Baxter Foundation.

Author Contributions A.S., H.M.B. and R.D. designed the research, and A.S. and R.D. performed the experiments with support from P.K. S.V. performed single-cell RT–PCR. A.S. analysed the data. A.S., R.D. and H.M.B. discussed the results and wrote the paper.

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Correspondence to Helen M. Blau.

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Sacco, A., Doyonnas, R., Kraft, P. et al. Self-renewal and expansion of single transplanted muscle stem cells. Nature 456, 502–506 (2008). https://doi.org/10.1038/nature07384

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