Nunclon delta surface plastic plates

Skeletal muscle-derived cells were differentiated in 24-well Nunclon™ Delta Surface plastic plates (Thermo Scientific, MA, USA) by replacing the growth medium with Skeletal Muscle Cell Differentiation Medium (500 ml, PromoCell GmbH, Germany), supplemented with 10 ml of Skeletal Muscle Cell Differentiation Medium Supplement Pack (PromoCell GmbH, Germany) and 240 μl gentamicin (8 mg/ml, Sandoz GmbH, Austria) as described before [29 (link)].

SMDCs were differentiated in 24-well Nunclon™ Delta Surface plastic plates (ThermoFisher Scientific, Roskilde, Denmark). Cells were seeded after coating the wells with 0.1% gelatin in 0.9% NaCl (CellGenix, Freiburg, Germany). The coating was performed by adding 300 μL of coating solution in each well and incubated overnight at 37°C. Afterwards, the coating solution was aspirated and cells were directly seeded in growth medium without any washing in a 24-well plate. Differentiation of mononucleated human myoblasts to syncytial myotubes was performed by replacing the growth medium with Skeletal Muscle Cell Differentiation medium (500 mL, PromoCell GmbH, Germany), supplemented with 10 mL of Skeletal Muscle Cell Differentation Medium Supplement Pack (PromoCell GmbH, Germany) and 240 μL gentamicin (8 mg/mL, Sandoz GmbH, Austria). Briefly, cells were seeded in a 24-well plate (120000 cells/per well) with growth medium for 1–2 days. For coating experiments, 200000 cells/well were seeded. Afterwards the growth medium was aspirated and cells were washed with Tyrode’s salt solution (Sigma-Aldrich Co. LLC, Germany). Finally, cells were covered with 1 mL of differentiation medium for 5–7 days without further medium change.