Directed Differentiation of Human Pluripotent Stem Cells into Skeletal Muscle Progenitor Cells

Human pluripotent stem cells (hPSCs) hold the potential to differentiate into skeletal muscle progenitor cells (SMPCs), which provide a human model for muscle disease modeling, drug screening, and a promising cell source for future stem cell-based muscle therapies. Directed differentiation is a transgene-free approach to guide hPSCs through the developmental path and differentiate them into embryonic-like SMPCs. However, the directed differentiation of functional SMPCs from hPSCs can be challenging due to cell line variations leading to low efficiency and reproducibility. In this section, we introduce a robust protocol that has been validated in multiple hPSC lines, providing cues on how to ensure efficient SMPC differentiation routinely. We describe the directed differentiation and isolation of SMPCs, using well-characterized markers ERBB3 and NGFR. Previous work from our lab has generated transcriptomic profiles of hPSC-derived SMPCs and functionally characterized the differentiated SMPCs in vitro and in vivo using different mouse models, and in this chapter, we will provide a detailed method for generating SMPCs used in these and additional studies.