文摘
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Summary
Human pluripotent stem cells (hPSCs) constitute a promising resource for use in cell-based therapies and a valuable in聽vitro model for聽studying early human development and disease. Despite significant advancements in the derivation of specific fates from hPSCs, the generation of skeletal muscle remains challenging and is mostly dependent on transgene expression. Here, we describe a method based on the use of a small-molecule GSK3尾 inhibitor to derive skeletal muscle from several hPSC lines. We show that early GSK3尾 inhibition is sufficient to create the conditions necessary for highly effective derivation of muscle cells. Moreover, we developed a strategy for stringent fluorescence-activated cell sorting-based purification of emerging PAX3+/PAX7+ muscle precursors that are able to differentiate in postsort cultures into mature myocytes. This transgene-free, efficient protocol provides an essential tool for producing myogenic cells for in聽vivo preclinical studies, in聽vitro screenings, and disease modeling.
