Validation of an in vitro muscle platform to evaluate myogenesis and calcium handling in control and dystrophic human myotubes

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dc.contributor.authorMosqueira Martín, Laura
dc.contributor.authorPrendes García, Carolina
dc.contributor.authorVesga Castro, Camila
dc.contributor.authorMarco Moreno, Pablo
dc.contributor.authorIrastorza Lorenzo, Ainhoa
dc.contributor.authorIzeta, Ander
dc.contributor.authorMadarieta, Iratxe
dc.contributor.authorMartí Carrera, Itxaso
dc.contributor.authorParedes, Jacobo
dc.contributor.authorLópez de Munain Arregui, Adolfo
dc.contributor.authorVallejo Illarramendi, Ainara
dc.date.accessioned2026-03-03T15:11:06Z
dc.date.available2026-03-03T15:11:06Z
dc.date.issued2026-12-01
dc.date.updated2026-03-03T15:11:06Z
dc.description.abstractElectrical impedance has emerged as a powerful tool for real-time, label-free, and non-invasive monitoring of cellular processes. Here, we employed an impedance-based assay to characterize the myogenic process of control and dystrophic human myoblasts. First, we conducted a comprehensive analysis of control myoblast differentiation, assessing the effects of initial seeding density and various extracellular matrix coatings. We also evaluated the influence of electrode presence and current application, both of which improved myoblast alignment. Immortalized myoblasts from Duchenne muscular dystrophy patients exhibited marked alterations in early differentiation and maturation, which were readily detected via impedance measurements. We further compared two differentiation protocols using one control and one dystrophic representative cell line. While both protocols supported the formation of mature myotubes, impedance profiles differed depending on the culture medium. Notably, we identified the protocol with superior impedance profile reproducibility over the culture lifespan. Finally, we successfully assessed calcium homeostasis in control and dystrophic myotubes differentiated on 96-well impedance plates. Our findings underscore the potential of impedance-based assays for monitoring myogenesis and identifying disease-associated phenotypes. Moreover, 96-well impedance plates represent a robust tool for high-throughput and high-content functional analysis in muscle disease modeling and therapeutic screening.en
dc.description.sponsorshipThis work was supported by Gobierno Vasco (KK-2019/00093, KK-2024/00111, 2022111014, 2022111045, 2024333014, MTVD22/BD/003, IT1732-22), Agencia Estatal de Investigación del Ministerio de Ciencia e Innovación funded by MICIU/AEI /10.13039/501100011033 (PID2020-119780RB-100; PID2024-157677OB-I00), and Diputación Foral de Gipuzkoa (2021-CIEN- 000043-01). L.M.-M. held a PhD fellowship from the UPV/EHU (PIF19/184), P.M.-M. held a PhD fellowship from Fundación Gangoiti Barrera, C.V.-C. held a PhD fellowship from Tecnun and is currently holding a Postdoctoral fellowship from the Basque Government (POS_2022_1_0042), and A. Irastorza held a PhD fellowship from the Basque Government (PRE_2019_1_0031)en
dc.identifier.citationMosqueira-Martín, L., Prendes-García, C., Vesga-Castro, C., Marco-Moreno, P., Irastorza, A., Izeta, A., Madarieta, I., Martí-Carrera, I., Paredes, J., de Munain, A. L., & Vallejo-Illarramendi, A. (2026). Validation of an in vitro muscle platform to evaluate myogenesis and calcium handling in control and dystrophic human myotubes. Scientific Reports, 16(1). https://doi.org/10.1038/S41598-025-31522-Z
dc.identifier.doi10.1038/S41598-025-31522-Z
dc.identifier.eissn2045-2322
dc.identifier.urihttps://hdl.handle.net/20.500.14454/5320
dc.language.isoeng
dc.publisherNature Research
dc.rights© The Author(s) 2025
dc.titleValidation of an in vitro muscle platform to evaluate myogenesis and calcium handling in control and dystrophic human myotubesen
dc.typejournal article
dcterms.accessRightsopen access
oaire.citation.issue1
oaire.citation.titleScientific Reports
oaire.citation.volume16
oaire.licenseConditionhttps://creativecommons.org/licenses/by/4.0/
oaire.versionVoR
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