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Nat Protoc. 2016 Oct;11(10):1833-50. doi: 10.1038/nprot.2016.110. Epub 2016 Sep 1.

Generation of human muscle fibers and satellite-like cells from human pluripotent stem cells in vitro.

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Institut de Génétique et de Biologie Moléculaireet Cellulaire (IGBMC), CNRS (UMR 7104), Inserm U964, Université de Strasbourg, Illkirch-Graffenstaden, France.
Department of Pathology, Brigham and Women's Hospital, Boston, Massachusetts, USA.
Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
Harvard Stem Cell Institute, Boston, Massachusetts, USA.
Anagenesis Biotechnologies, Parc d'innovation, Illkirch-Graffenstaden, France.
Disease Biophysics Group, Wyss Institute for Biologically Inspired Engineering, School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts, USA.


Progress toward finding a cure for muscle diseases has been slow because of the absence of relevant cellular models and the lack of a reliable source of muscle progenitors for biomedical investigation. Here we report an optimized serum-free differentiation protocol to efficiently produce striated, millimeter-long muscle fibers together with satellite-like cells from human pluripotent stem cells (hPSCs) in vitro. By mimicking key signaling events leading to muscle formation in the embryo, in particular the dual modulation of Wnt and bone morphogenetic protein (BMP) pathway signaling, this directed differentiation protocol avoids the requirement for genetic modifications or cell sorting. Robust myogenesis can be achieved in vitro within 1 month by personnel experienced in hPSC culture. The differentiating culture can be subcultured to produce large amounts of myogenic progenitors amenable to numerous downstream applications. Beyond the study of myogenesis, this differentiation method offers an attractive platform for the development of relevant in vitro models of muscle dystrophies and drug screening strategies, as well as providing a source of cells for tissue engineering and cell therapy approaches.

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