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.

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.

Department of Developmental Biology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, 75390-9133, Texas, USA

Radbod Darabi, Kimberly Gehlbach, Shwetha Kamath, Mitsujiro Osawa, Michael Kyba & Rita C R Perlingeiro

Department of Neurology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, 75390-9133, Texas, USA

Department of Physiology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, 75390-9133, Texas, USA

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.

Supplementary Figures 1–6, Supplementary Table 1 and Supplementary Methods (PDF 928 kb)

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