Electrical Pulse Stimulation of Primary Human Skeletal Muscle Cells
Electrical pulse stimulation (EPS) is an in vitro method of inducing contractions in cultured skeletal muscle cells of human and animal origin. Motor neuron activation of muscle fibers can be replaced by applying EPS on differentiated skeletal muscle cells (myotubes) in culture (Thelen et al. Biochemical J 321:845–848, 1997, Fujita et al. Exp Cell Res 313:1853–1865, 2007).
Here we describe two protocols for EPS of human myotubes in 6-well plates: acute, high-frequency (single bipolar pulses of 2 ms, 100 Hz for 200 ms every fifth second for 5–60 min, 10–30 V) and chronic, low-frequency (single bipolar pulses of 2 ms, 1 Hz 10–30 V for 48 h) at the end of a 7 days long differentiation.
Key wordsSkeletal muscle Myotubes Contractions Electrical pulse stimulations
- 3.Askanas V, Kwan H, Alvarez RB, Engel WK, Kobayashi T, Martinuzzi A, Hawkins EF (1987) De novo neuromuscular junction formation on human muscle fibres cultured in monolayer and innervated by foetal rat spinal cord: ultrastructural and ultrastructural--cytochemical studies. J Neurocytol 16(4):523–537CrossRefGoogle Scholar
- 6.Nikolic N, Bakke SS, Kase ET, Rudberg I, Flo Halle I, Rustan AC, Thoresen GH, Aas V (2012) Electrical pulse stimulation of cultured human skeletal muscle cells as an in vitro model of exercise. PLoS One 7(3):e33203. https://doi.org/10.1371/journal.pone.0033203CrossRefPubMedPubMedCentralGoogle Scholar
- 7.Feng YZ, Nikolic N, Bakke SS, Kase ET, Guderud K, Hjelmesaeth J, Aas V, Rustan AC, Thoresen GH (2015) Myotubes from lean and severely obese subjects with and without type 2 diabetes respond differently to an in vitro model of exercise. Am J Physiol Cell Physiol 308(7):C548–C556. https://doi.org/10.1152/ajpcell.00314.2014CrossRefPubMedGoogle Scholar
- 9.Lambernd S, Taube A, Schober A, Platzbecker B, Gorgens SW, Schlich R, Jeruschke K, Weiss J, Eckardt K, Eckel J (2012) Contractile activity of human skeletal muscle cells prevents insulin resistance by inhibiting pro-inflammatory signalling pathways. Diabetologia 55(4):1128–1139. https://doi.org/10.1007/s00125-012-2454-zCrossRefPubMedGoogle Scholar