Muscle satellite cells are specific stem cells of skeletal muscle. They play important roles in muscle growth, hypertrophy, and regeneration. The transcriptional factor FOXO1, known to induce muscle atrophy, has been reported to be involved in muscle differentiation; however, the experiments have been performed using the immortalized cell line C2C12. In this study, we evaluated the differentiation capacity of satellite cells isolated from muscle‐specific FOXO1‐transgenic (Tg) mice because these cells are believed to mimic in vivo skeletal muscles. Extensor digitorum longus muscles were isolated from mice and digested in collagenase‐type 1 solution until they separated into single fibers. The single viable muscle fibers were then purified using Pasteur pipettes with fire‐polished tips and cultured in growth medium in Matrigel‐coated wells. After cultivation for 5 days in growth medium, followed by 3 days in differentiation medium, we evaluated the differentiation capacity of the satellite cells. Thus, satellite cells were successfully isolated from FOXO1‐Tg mice. These cells tended to exhibit suppressed myotube formation compared with those from the wild‐type mice. These findings suggested that FOXO1 plays a role in not only mature skeletal muscles but also muscle satellite cells.
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