Functional skeletal muscle regeneration from differentiating embryonic stem cells


Little progress has been made toward the use of embryonic stem (ES) cells to study and isolate skeletal muscle progenitors. This is due to the paucity of paraxial mesoderm formation during embryoid body (EB) in vitro differentiation and to the lack of reliable identification and isolation criteria for skeletal muscle precursors. Here we show that expression of the transcription factor Pax3 during embryoid body differentiation enhances both paraxial mesoderm formation and the myogenic potential of the cells within this population. Transplantation of Pax3-induced cells results in teratomas, however, indicating the presence of residual undifferentiated cells. By sorting for the PDGF-α receptor, a marker of paraxial mesoderm, and for the absence of Flk-1, a marker of lateral plate mesoderm, we derive a cell population from differentiating ES cell cultures that has substantial muscle regeneration potential. Intramuscular and systemic transplantation of these cells into dystrophic mice results in extensive engraftment of adult myofibers with enhanced contractile function without the formation of teratomas. These data demonstrate the therapeutic potential of ES cells in muscular dystrophy.

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Figure 1: Characterization of iPax3 ES cells.
Figure 2: Transplantation of unpurified iPax3 EB-derived cells into cardiotoxin-injured Rag2−/−γc−/− mice.
Figure 3: Effects of Pax3 induction on paraxial mesoderm formation during EB development.
Figure 4: Characterization of Pax3-induced PDGF-αR+Flk-1 derived cells.
Figure 5: Robust engraftment on transplantation of Pax3-induced PDGFαR+Flk1 EB-derived cells into cardiotoxin-injured Rag2−/−γc−/− and dystrophic mice.
Figure 6: Systemic cell transplantation improves contractile properties of tibialis anterior muscles in mdx mice.


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We thank J. Stull for discussion and M. Rudnicki for technical advice. The monoclonal antibody to MHC was obtained from the Developmental Studies Hybridoma Bank, which was developed under the auspices of the National Institute of Child Health and Human Development and is maintained by the University of Iowa. This work was supported by the Dr. Bob and Jean Smith Foundation.

Author information




R.D. designed and conducted the in vitro and in vivo experiments with iPax3 ES cells. R.D. also performed the final analysis of the data and contributed to writing the paper. K.G. generated the iPax3 ES cell line. R.M.B. performed the i.a. transplantations. S.K. performed quantifications for dystrophin and assisted R.D. with some of the staining. M.O. performed the i.v. transplantations. K.E.K. supervised and assisted on interpretation of the muscle functional analyses. M.K. provided materials, supervised K.G. on generating the cell line and analyzed the data. R.C.R.P. supervised the overall project, designed experiments, analyzed the data and wrote the paper.

Corresponding author

Correspondence to Rita C R Perlingeiro.

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Darabi, R., Gehlbach, K., Bachoo, R. et al. Functional skeletal muscle regeneration from differentiating embryonic stem cells. Nat Med 14, 134–143 (2008).

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