Isolation of human fibroadipogenic progenitors and satellite cells from frozen muscle biopsies
Xavier Suárez‐Calvet, Esther Fernández‐Simón, Patricia Piñol‐Jurado, Jorge Alonso‐Pérez, Ana Carrasco‐Rozas, Cinta Lleixà, Susana López‐Fernández, Gemma Pons, Laura Soria, Anne Bigot, Vincent Mouly, Isabel Illa, Eduard Gallardo, Jyoti K. Jaiswal, J. Diaz-Manera
Abstract
Skeletal muscle contains multiple cell types that work together to maintain tissue homeostasis. Among these, satellite cells (SC) and fibroadipogenic progenitors cells (FAPs) are the two main stem cell pools. Studies of these cells using animal models have shown the importance of interactions between these cells in repair of healthy muscle, and degeneration of dystrophic muscle. Due to the unavailability of fresh patient muscle biopsies, similar analysis of interactions between human FAPs and SCs is limited especially among the muscular dystrophy patients. To address this issue here we describe a method that allows the use of frozen human skeletal muscle biopsies to simultaneously isolate and grow SCs and FAPs from healthy or dystrophic patients. We show that while the purified SCs differentiate into mature myotubes, purified FAPs can differentiate into adipocytes or fibroblasts demonstrating their multipotency. We find that these FAPs can be immortalized and the immortalized FAPs (iFAPs) retain their multipotency. These approaches open the door for carrying out personalized analysis of patient FAPs and interactions with the SCs that lead to muscle loss.