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. 2022 Jan 17;12:827. doi: 10.1038/s41598-022-04804-z

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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The experimental schematic workflow in this study. C2C12 cells were cultured for 7 days in the following 4 different types of mediums. (i) DMEM; only basic medium without any supplementation, (ii) Serum; DMEM supplemented with animal-derived serum. 10% of fetal bovine serum (FBS) and 2% of horse serum (HS) were used for proliferation and differentiation, respectively. (iii) B27; chemically defined serum substitute, 2% and 1% of B27 were supplemented in DMEM for proliferation and differentiation, respectively. (iv) AIM-V, chemically defined medium. Cell morphology and viability were monitored for the entire culture period. Cell growth was further profiled during the proliferation status, CK activity, myotube diameter, muscle contraction were further assessed for myogenic differentiation. Finally, culture samples from day 1 and day 7 representing myoblasts and myotubes were analyzed for the metabolomic study.