PT  - JOURNAL ARTICLE
AU  - Travis Ruan
AU  - Dylan Harney
AU  - Yen Chin Koay
AU  - Lipin Loo
AU  - Mark Larance
AU  - Leslie Caron
TI  - Anabolic Factors and Myokines Improve Differentiation of Human Embryonic Stem Cell Derived Skeletal Muscle Cells
AID  - 10.1101/2022.02.08.477036
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.02.08.477036
4099  - http://biorxiv.org/content/early/2022/02/08/2022.02.08.477036.short
4100  - http://biorxiv.org/content/early/2022/02/08/2022.02.08.477036.full
AB  - Skeletal muscle weakness is linked to many adverse health outcomes. Current research to identify new drugs has often been inconclusive due to lack of adequate cellular models. We have previously developed a scalable monolayer system to differentiate human embryonic stem cell (hESC) into mature skeletal muscle cells (SkMC) within 26 days without cell sorting or genetic manipulation. Here, building on our previous work, we show that differentiation and fusion of myotubes can be further enhanced using the anabolic factors testosterone (T) and follistatin (F) in combination with a cocktail of myokines (C). Importantly, combined TFC treatment significantly enhanced both hESC-SkMC fusion index and expression of various skeletal muscle markers including the motor protein Myosin Heavy Chain (MyHC). Transcriptomic and proteomic analysis revealed oxidative phosphorylation as the most up-regulated pathway and a significantly higher level of ATP and increased mitochondrial mass were also observed in TFC-treated hESC-SkMCs, suggesting enhanced energy metabolism is coupled to improved muscle differentiation. This cellular model will be a powerful tool for studying in vitro myogenesis and for drug discovery to further enhance muscle development or treat muscle diseases.Competing Interest StatementThe authors have declared no competing interest.hESChuman embryonic stem cellSkMCskeletal muscle cellsTtestosteroneFfollistatinCcocktail of myokinesTFCtestosterone follistatin and cocktail of myokinesMyHCmyosin heavy chainhPSChuman pluripotent stem cellshiPSChuman induced pluripotent stem cellsMRFmyogenic regulatory factorsIL4interleukin-4IL6interleukin-6BDNFbrain derived neurotrophic factorVEGFvascular endothelial growth factorNTCnon-treated cells