{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T14:47:04Z","timestamp":1786978024744,"version":"build-2736575974"},"reference-count":44,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2025,12,4]],"date-time":"2025-12-04T00:00:00Z","timestamp":1764806400000},"content-version":"vor","delay-in-days":3,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,12,1]],"date-time":"2025-12-01T00:00:00Z","timestamp":1764547200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32402710"],"award-info":[{"award-number":["32402710"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J cachexia sarcopenia muscle"],"published-print":{"date-parts":[[2025,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Apoptosis coincides with the differentiation of skeletal myoblasts, and numerous studies have shown that the apoptotic activity is required for myogenic differentiation. Although the role of apoptosis in skeletal muscle differentiation has been well documented, its mechanism is largely unknown.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Apoptotic extracellular vesicles (apoVs) were extracted from differentiated C2C12 cells or STS\u2010treated undifferentiated C2C12 myoblasts. C2C12 myoblasts, 8\u2010week\u2010old male mice or 15\u2010month\u2010old male mice were used as in\u00a0vitro and in\u00a0vivo models, respectively. These models were treated with C2C12\u2010derived apoVs to explore the biological function and mechanism of apoVs in myogenic differentiation, skeletal muscle development and aging.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Proteomic analysis revealed that inhibition of apoptotic activity by Z\u2010VAD\u2010FMK (ZVAD) affected extracellular components. Using immunofluorescence staining, western blotting and transmission electron microscopy analysis, our results demonstrated the generation of apoVs during myogenic differentiation. C2C12\u2010derived apoVs exhibited a typical double\u2010membrane spherical structure, phosphatidylserine exposure and were highly positive for the general apoV markers cleaved caspase 3 (CASP3), Alix and TSG101. Inhibition of apoptotic activity significantly reduced (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009=\u20090.0029) the protein level of myosin heavy chain (1.05 in the Con group vs. 0.38 in the ZVAD group) accompanied by a marked decrease (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.0001) in apoV production (5.91e+10\u2009\u00b1\u20098.93e+09 in the Con group vs. 1.77e+10\u2009\u00b1\u20091.36e+09 in the ZVAD group). Proteomic analysis of apoVs suggested that C2C12\u2010derived apoVs contain multiple pro\u2010differentiation proteins, including insulin\u2010like growth factor 1 receptor (Igf1r). ApoVs could be taken up by recipient cells and subsequently rescued the impaired C2C12 differentiation induced by ZVAD (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20090.0001) and promoted the normal myogenic differentiation process (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009=\u20090.0081) by carrying Igf1r and promoting PI3K\/AKT\/mTOR activation. Knockdown of Igf1r or inhibition of PI3K activation diminished the positive role of apoVs. In addition, apoV treatment promoted skeletal muscle development in 8\u2010week\u2010old male mice (\n                      <jats:italic>n<\/jats:italic>\n                      \u2009=\u20096, Cohen's d\u2009=\u20091.993, power\u2009=\u20090.874) and relieved age\u2010related muscle loss (\n                      <jats:italic>n<\/jats:italic>\n                      \u2009=\u20096, Cohen's d\u2009=\u20093.97, power\u2009=\u20090.999).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>In summary, this study demonstrates the generation of apoVs during myogenic differentiation. The apoVs derived from skeletal muscle cells promote skeletal muscle cell differentiation and delay age\u2010related muscle loss. These results provide a theoretical basis for elucidating the mechanism of skeletal muscle development and treating skeletal muscle\u2010related diseases.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/jcsm.70159","type":"journal-article","created":{"date-parts":[[2025,12,5]],"date-time":"2025-12-05T02:50:29Z","timestamp":1764903029000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["C2C12\u2010Derived ApoVs Promote Skeletal Muscle Development and Ameliorate Age\u2010Related Muscle Loss Through Igf1r\/PI3K\/AKT\/mTOR Pathway"],"prefix":"10.1002","volume":"16","author":[{"given":"Aiwen","family":"Jiang","sequence":"first","affiliation":[{"name":"Key Laboratory for Animal Genetics, Breeding, Reproduction and Molecular Design, College of Animal Science and Technology Yangzhou University  Yangzhou 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