MilliporeSigma
  • Home
  • Search Results
  • Generation of human muscle fibers and satellite-like cells from human pluripotent stem cells in vitro.

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

Nature protocols (2016-09-02)
Jérome Chal, Ziad Al Tanoury, Marie Hestin, Bénédicte Gobert, Suvi Aivio, Aurore Hick, Thomas Cherrier, Alexander P Nesmith, Kevin K Parker, Olivier Pourquié
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
TWEEN® 20, BioXtra, viscous liquid
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, pH 7, ≥98%
Sigma-Aldrich
Monoclonal Anti-Myosin (Skeletal, Fast) antibody produced in mouse, clone MY-32, ascites fluid
Sigma-Aldrich
Venor GeM Mycoplasma Detection Kit, PCR-based
Sigma-Aldrich
Trypan Blue solution, 0.4%, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Triton X-100, for molecular biology
Sigma-Aldrich
Dimethyl sulfoxide, Hybri-Max, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%