Abstract
The plasticizer di (2-ethylhexyl) phthalate (DEHP) inhibits differentiation, impairs glucose metabolism, and decreases mitochondrial function in murine muscle satellite cells; however, if these effects are translated to human cells is unknown. The goal of this study was to evaluate changes in morphology and proliferation of primary human skeletal muscle cells exposed to DEHP. Rectus abdominis muscle samples were obtained from healthy women undergoing programed cesarean surgery. Skeletal muscle cells were isolated and grown under standard primary culture conditions, generating two independent sample groups of 25 subcultures each. Cells from the first group were exposed to 1 mM DEHP for 13 days and monitored for changes in cell morphology, satellite cell frequency and total cell abundance, while the second group remained untreated (control). Differences between treated and untreated groups were compared using generalized linear mixed models (GLMM). Cell membrane and nuclear envelope boundary alterations, loss of cell volume and presence of stress bodies were observed in DEHP-treated cultures. DEHP-treated cultures also showed a significant reduction in satellite cell frequency compared to controls. Exposure to DEHP reduced human skeletal muscle cell abundance. Statistical differences were found between the GLMM slopes, suggesting that exposure to DEHP reduced growth rate. These results suggest that exposure to DEHP inhibits human skeletal muscle cell proliferation, as evidenced by reduced cell abundance, potentially compromising long-term culture viability. Therefore, DEHP induces human skeletal muscle cell deterioration potentially inducing an inhibitory effect of myogenesis by depleting satellite cells.
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The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
Financial support for this study was provided by Consejo Nacional de Ciencia y Tecnología (CONACYT; Fronteras 2013-01-2305). EBP is a recipient of a graduate scholarship from CONACYT (# 613207). Authors are members of the CYTED network RIESCOS (ref. 419RT0578). All samples were collected in accordance with the guidelines of Mexico’s Comisión Nacional de Bioética (CONBIOÉTICA) and the Code of Ethics of the World Medical Association (Declaration of Helsinki). The research protocol and informed consent forms were registered and approved by Comité de Ética en Investigación and Comité Hospitalario de Bioética (F-CNIC 2019-174 and R 2000-785-008), Comisión Nacional de Investigación Científica del Instituto Mexicano del Seguro Social (IMSS; 2018-785-010), as well as by CONBIOÉTICA (09-CEI-009-20160601). Authors acknowledge personnel at IMSS for their help in sample collection, and O. Lugo-Lugo and P. Hernandez-Almaraz for their technical support and assistance during this study.
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Financial support for this study was provided by Consejo Nacional de Ciencia y Tecnología (CONACYT; Fronteras 2013-01-2305).
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EB-P, sample collection and processing, bioassays, data collection and processing, manuscript production; VL-M, data processing, statistical analyses, manuscript edition; CH-C, sample collection, data analyses, manuscript edition; RG-R, sample collection, data analyses, manuscript edition; JPV-M, data analyses, manuscript edition; TZ-S, experimental design, sample collection and processing, bioassays, data processing, manuscript edition.
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Brassea-Pérez, E., Labrada-Martagón, V., Hernández-Camacho, C.J. et al. DEHP exposure impairs human skeletal muscle cell proliferation in primary culture conditions: preliminary study. Cytotechnology 75, 335–348 (2023). https://doi.org/10.1007/s10616-023-00580-4
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DOI: https://doi.org/10.1007/s10616-023-00580-4