Dispase/Collagenase cocktail allows for co-isolation of satellite cells and fibro-adipogenic progenitors from human skeletal muscle.

Satellite cells (SCs) and fibro-adipogenic progenitors (FAPs) are progenitor populations found in muscle that form new myofibers post-injury. Muscle development, regeneration, and tissue engineering experiments require robust progenitor populations, yet their isolation and expansion are difficult given their scarcity in muscle, limited muscle biopsy sizes in humans, and lack of methodological detail in the literature. Here, we investigated whether a dispase and collagenase type 1 & 2 cocktail could allow dual isolation of SCs and FAPs, enabling significantly increased yield from human skeletal muscle. Post-dissociation, we found that single cells could be sorted into CD56+CD31-CD45- (SC) and CD56-CD31-CD45- (FAP) cell populations, expanded in culture, and characterized for lineage-specific marker expression and differentiation capacity; we obtained ~10% SCs and ~40% FAPs, with yields 2-fold better than what is reported in current literature. SCs were PAX7+ and retained CD56 expression and myogenic fusion potential after multiple passages, expanding up to 10