Sphingosine-1-phosphate pretreatment amends hypoxia-induced metabolic dysfunction and impairment of myogenic potential in differentiating C2C12 myoblasts by stimulating viability, calcium homeostasis and energy generation

Abstract

Sphingosine-1-phosphate (S1P) has a role in transpiration in patho-physiological signaling in skeletal muscles. The present study evaluated the pre-conditioning efficacy of S1P in facilitating differentiation of C2C12 myoblasts under a normoxic/hypoxic cell culture environment. Under normoxia, exogenous S1P significantly promoted C2C12 differentiation as evident from morphometric descriptors and differentiation markers of the mature myotubes, but it could facilitate only partial recovery from hypoxia-induced compromised differentiation. Pretreatment of S1P optimized the myokine secretion, intracellular calcium release and energy generation by boosting the aerobic/anaerobic metabolism and mitochondrial mass. In the hypoxia-exposed cells, there was derangement of the S1PR expression patterns, while the same could be largely restored with S1P pretreatment. This is being proposed as a plausible underlying mechanism for the observed pro-myogenic efficacy of exogenous S1P preconditioning. The present findings are an invaluable addition to the existing knowledge on the pro-myogenic potential of S1P and may prove beneficial in the field of hypoxia-related myo-pathologies.

The secretome of human dental pulp stem cells protects myoblasts from hypoxia‑induced injury via the Wnt/β‑catenin pathway

“…Previous studies have demonstrated pathological hypoxia-induced skeletal muscle myoblast injury, including the inhibition of viability and differentiation (43)(44)(45). Regarding viability, cocl 2 produced changes typical of cell death, such as characteristic cell extension, increased volume, chromatin condensation and G2/M phase cell cycle arrest, ultimately leading to reduced proliferation (13).…”

Automatic Quantitative Segmentation of Myotubes Reveals Single-cell Dynamics of S6 Kinase Activation

“…Therefore, proper background correction is needed for quantitative signal acquisi- tion (Zimmermann et al, 2003). Manual selection of background regions near the regions of interest (ROIs) has been used widely for background correction (Ceelen et al, 2007;Horie et al, 2015;Rahar et al, 2018). However, in the case of differentiated C2C12 cells, it is difficult to manually identify background regions, because reserve cells occupy spaces between myotubes ( Fig.…”

“…developed. Indeed, intracellular signal activity and reactive oxygen species in myotubes have been measured in vitro only by manual ROI selection of myotubes (Ceelen et al, 2007;Horie et al, 2015;Rahar et al, 2018). Since our method does not make assumptions on the number of nuclei and cell shape, our method can be used for any type of polynuclear and deformed cells.…”

“…C2C12 cells, derived from mice myoblasts, have been used as model cell lines for the study of cell differentiation and muscle functions in vitro (Fodor et al, 2017;Rahar et al, 2018;Rapizzi et al, 2008;Soltow et al, 2013). By differentiation induction of C2C12 cells, a large subpopulation of myoblasts forms elongated polynuclear myotubes, while a subpopulation of the cells, called reserve cells, remains undifferentiated ( Fig.…”

Biomaterials-Induced Stem Cells Specific Differentiation Into Intervertebral Disc Lineage Cells

“…Moreover, a few studies have investigated the effect of hypoxia on AF and NP cells seeded on 3D scaffolds and found that hypoxia was effective in maintaining the NP cell phenotype but does not affect AF cells (Mwale et al, 2011;Feng et al, 2013). Therefore, future development of IVD biomaterials should take oxygen content into account and use chemicals (CoCl 2 et al) or other methods to induce cellular hypoxia response, so as to improve efficiency of inducing tissue specific differentiation, while minimize the unfavorable situation on cells survival induced by prolonged hypoxia (Rahar et al, 2018).…”