Mild Hyperthermia-Induced Myogenic Differentiation in Skeletal Muscle Cells: Implications for Local Hyperthermic Therapy for Skeletal Muscle Injury
Mild heat treatment promotes myogenic differentiation in a treatment time-dependent manner. (a) Myoblasts were differentiated in DMEM supplemented with 2% horse serum under specified temperature condition for 5 days. Differentiating myoblasts were observed and counted every 24 hours under phase contrast microscope (magnification = 10x). Scale bar represents 100 μm. (b) Myotubes were counted and expressed as number per field. (c) Differentiated myotubes were stained with DAPI and MF20 antibody and were visualized by fluorescence microscope (magnification = 20x). Scale bar represents 50 μm. (d) Myonuclei, (e) fusion index, (f) myotube width, (g) and myotube length were quantified. Each experiment was performed with at least four separate cell cultures. Bar graph represented means ± SEM. The data were analyzed using one-way ANOVA followed by Tukey’s post hoc test. versus NT (no treatment); versus IHT (intermittent heat treatment).