The effect of differentiation and TGFβ on mitochondrial respiration and mitochondrial enzyme abundance in cultured primary human skeletal muscle cells
Autor: | Lisa Kappler, Christoph Hoffmann, Hans-Ulrich Häring, Selina Höckele, Martin Hrabĕ de Angelis, Cora Weigert |
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Rok vydání: | 2017 |
Předmět: |
0301 basic medicine
Male Cellular respiration Cellular differentiation Cell Respiration lcsh:Medicine Oxidative phosphorylation Mitochondrion Article Electron Transport Electron Transport Complex IV 03 medical and health sciences 0302 clinical medicine Transforming Growth Factor beta medicine Myocyte Humans lcsh:Science Muscle Skeletal Cells Cultured Muscle Cells Multidisciplinary Myogenesis Muscle cell differentiation Chemistry lcsh:R Skeletal muscle Cell Differentiation Middle Aged Cell biology Mitochondria Oxygen 030104 developmental biology medicine.anatomical_structure lcsh:Q 030217 neurology & neurosurgery |
Zdroj: | Scientific Reports Sci. Rep. 8:737 (2018) Scientific Reports, Vol 8, Iss 1, Pp 1-12 (2018) |
ISSN: | 2045-2322 |
Popis: | Measuring mitochondrial respiration in cultured cells is a valuable tool to investigate the influence of physiological and disease-related factors on cellular metabolism; however, the details of the experimental workflow greatly influence the informative value of the results. Working with primary cells and cell types capable of differentiation can be particularly challenging. We present a streamlined workflow optimised for investigation of primary human skeletal muscle cells. We applied the workflow to differentiated and undifferentiated cells and we investigated the effect of TGFβ1 treatment. Differentiation of myoblasts to myotubes increased mitochondrial respiration and abundance of mitochondrial enzymes and mitochondrial marker proteins. Differentiation also induced qualitative changes in mitochondrial protein composition and respiration. TGFβ1 reduced complex IV protein MTCO1 abundance in both myoblasts and myotubes. In myoblasts, spare electron transport system (ETS) capacity was reduced due to a reduction in maximal oxygen consumption. In TGFβ1-treated myotubes, the reduction in spare ETS capacity is mainly a consequence of increased oxidative phosphorylation capacity and complex III protein UQCRC2. Taken together, our data shows that it is important to monitor muscle cell differentiation when mitochondrial function is studied. Our workflow is not only sensitive enough to detect physiological-sized differences, but also adequate to form mechanistic hypotheses. |
Databáze: | OpenAIRE |
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