Real-time PCR analysis of microRNA expression in differentiating human skeletal muscle cells

Human skeletal myoblast cell line LHCN-M2 (Zhu et al., Aging Cell (2007), vol. 6, pp 515-523) were plated on collagen-coated 6-well Falcon cell culture plates at 500 000 cells/well, differentiation was induced 24h later by switching to DMEM supplemented with 0,01 mg/ml insulin and 0,1 mg/ml transferrin. Cells were collected at 0, 3, and 7 days of differentiation time points. Total RNA was extracted and miRNA expression evaluated by quantitative real-time PCR. MiRNA expression profiling in proliferating and differentiating (3 and 7 days) human skeletal myoblasts LHCN-M2.