TY - JOUR AU - Le Gall, Laura AU - Ouandaogo, Zamalou Gisele AU - Anakor, Ekene AU - Connolly, Owen AU - Butler Browne, Gillian AU - Laine, Jeanne AU - Duddy, William AU - Duguez, Stephanie PY - 2020 DA - 2020/07/08 TI - Optimized method for extraction of exosomes from human primary muscle cells JO - Skeletal Muscle SP - 20 VL - 10 IS - 1 AB - Skeletal muscle is increasingly considered an endocrine organ secreting myokines and extracellular vesicles (exosomes and microvesicles), which can affect physiological changes with an impact on different pathological conditions, including regenerative processes, aging, and myopathies. Primary human myoblasts are an essential tool to study the muscle vesicle secretome. Since their differentiation in conditioned media does not induce any signs of cell death or cell stress, artefactual effects from those processes are unlikely. However, adult human primary myoblasts senesce in long-term tissue culture, so a major technical challenge is posed by the need to avoid artefactual effects resulting from pre-senescent changes. Since these cells should be studied within a strictly controlled pre-senescent division count (<21 divisions), and yields of myoblasts per muscle biopsy are low, it is difficult or impossible to amplify sufficiently large cell numbers (some 250 × 106 myoblasts) to obtain sufficient conditioned medium for the standard ultracentrifugation approach to exosome isolation. SN - 2044-5040 UR - https://doi.org/10.1186/s13395-020-00238-1 DO - 10.1186/s13395-020-00238-1 ID - Le Gall2020 ER -