We show that CHIR can mediate efficient skeletal myogenesis

We show that SNDX-275 cost CHIR can mediate efficient skeletal myogenesis in mESCs using a 2 day CHIR treatment similar to that used for hESCs, illustrating the effectiveness of our protocol across these species. Our current findings are consistent with our previous results showing that Wnt signaling via CTNNB1 is important for murine P19 embryonal carcinoma cell differentiation, using gain- and loss-of-function as well as a dominant-negative approach (Petropoulos and Skerjanc, 2002; Wong et al., 2013). In addition, the greater experimental variability with mouse compared to human ESC differentiation into muscle could be due to the EB-based nature of the mESC serum-free protocol. Therefore, we are adapting the more consistent monolayer approach used with our hESC method to mESCs. Although hESCs are more therapeutically relevant, mESCs continue to play an integral role in our ability to study cellular differentiation, primarily due to their substantially shorter differentiation time, creating a simpler high-throughput model system for detailed molecular analysis. Overall, the results presented here have provided a method to differentiate mouse and human ESCs into highly enriched skeletal muscle lineage cultures. Most importantly, we show a stable PAX7+ve population of MPCs even after 7 weeks of culture. Ultimately, these studies provide the foundation for in vitro study of ESC skeletal myogenesis, which could contribute to future drug testing or stem cell therapies, leading to the repair of muscle in patients with muscular wasting disorders.
Experimental Procedures
Acknowledgments