Conditionally immortalised equine skeletal muscle cell lines for in vitro analysis.
| Autor: | Rooney MF; School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute (TBSI), Trinity College Dublin, 152-160, Pearse Street, Dublin 2, Ireland., Neto NGB; Trinity Centre for Biomedical Engineering, Trinity Biomedical Sciences Institute (TBSI), Trinity College Dublin, 152-160, Pearse Street, Dublin 2, Ireland.; Department of Mechanical, Manufacturing and Biomedical Engineering, Trinity College Dublin, Dublin 2, Ireland., Monaghan MG; Trinity Centre for Biomedical Engineering, Trinity Biomedical Sciences Institute (TBSI), Trinity College Dublin, 152-160, Pearse Street, Dublin 2, Ireland.; Department of Mechanical, Manufacturing and Biomedical Engineering, Trinity College Dublin, Dublin 2, Ireland., Hill EW; Plusvital Ltd., The Highline, Pottery Road, Dun Laoghaire, Co. Dublin, Ireland.; School of Agriculture and Food Science, University College Dublin, Belfield, Dublin 4, Ireland., Porter RK; School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute (TBSI), Trinity College Dublin, 152-160, Pearse Street, Dublin 2, Ireland. |
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| Jazyk: | angličtina |
| Zdroj: | Biochemistry and biophysics reports [Biochem Biophys Rep] 2022 Dec 05; Vol. 33, pp. 101391. Date of Electronic Publication: 2022 Dec 05 (Print Publication: 2023). |
| DOI: | 10.1016/j.bbrep.2022.101391 |
| Abstrakt: | Background: Thoroughbred racehorse performance is largely influenced by a major quantitative trait locus at the myostatin ( MSTN ) gene which determines aptitude for certain race distances due to a promoter region insertion mutation influencing functional phenotypes in skeletal muscle. To develop an in vitro system for functional experiments we established three novel equine skeletal muscle cell lines reflecting the variation in phenotype associated with MSTN genotype (CC/II, CT/IN and TT/NN for SNP g.66493737C > T/SINE insertion 227 bp polymorphism). Primary equine skeletal muscle myoblasts, isolated from Thoroughbred horse gluteus medius , were conditionally immortalised and evaluated to determine whether cell phenotype and metabolic function were comparable to functional characteristics previously reported for ex vivo skeletal muscle isolated from Thoroughbred horses with each genotype. Results: Primary myoblasts conditionally immortalised with the temperature sensitive SV40TtsA58 lentivirus vector successfully proliferated and could revert to their primary cell phenotype and differentiate into multinucleated myotubes. Skeletal muscle fibre type, MSTN gene expression, mitochondrial abundance, and mitochondrial function of the three MSTN genotype cell lines, were consistent with equivalent characterisation of ex vivo skeletal muscle samples with these genotypes. Furthermore, addition of coenzyme Q Conclusions: The observation that the phenotypic characteristics and metabolic function of the cells lines are equivalent to ex vivo skeletal muscle indicates that this in vitro system will enable efficient and cost-effective analyses of equine skeletal muscle for a range of different applications including understanding metabolic function, testing of nutritional supplements, drug test development and gene doping test development. In the multi-billion-euro international Thoroughbred horse industry research advances in the biological function of skeletal muscle are likely to have considerable impact. Furthermore, this novel genotype-specific system may be adapted and applied to human biomedicine to improve understanding of the effects of myostatin in human physiology and medicine. Competing Interests: EWH is a shareholder in and Chief Scientific Officer of Plusvital Ltd., an equine nutrition and genetic testing company. Plusvital Ltd. has been granted a licence for commercial use of the data that is contained within multiple granted patents and patent applications including (patent reference numbers): EP2352850, JP5667057, US8771943, AU2009290452, NZ591711, US9249470 and US2016215335. EWH is named on these patents. MFR, NGBN, MGM and RKP declare that they have no competing interests. Other than EWH, Plusvital Ltd. played no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. (© 2022 The Authors.) |
| Databáze: | MEDLINE |
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