DF-1 cells culture: DF-1 cell line of chicken embryo fibroblast were cultured in DMEM (Gibco, USA) supplemented with 10% (v/v) fetal bovine serum (FBS, Hyclone, USA) and 0.2% penicillin/streptomycin (Invitrogen, USA).
QM-7 cell culture: QM-7 cell lines of avian myogenic origin were cultured in high-glucose M199 medium (Gibco, USA) with 10% fetal bovine serum (FBS), 10% tryptose phosphate broth solution (Sigma, USA), and 0.2% penicillin/streptomycin.
CPM isolation and culture: Primary myoblasts were isolated from the leg muscle of 11-day old chicken embryos. First, the muscle tissues were dissected away from the skin and bone, and then homogenized in a petri dish. To release single cells, the suspension was digested with pancreatin for 20 min at 37 °C. After neutralization with complete medium, single cells were collected by centrifugation at 500 × g. Subsequently, serial plating was performed to enrich for primary myoblasts and eliminate fibroblasts. Primary myoblasts were cultured in Roswell Park Memorial Institute (RPMI)-1640 medium (Gibco, USA) with 20% FBS, 1% nonessential amino acids, and 0.2% penicillin/streptomycin. The differentiation of myoblasts was induced by RPMI-1640 medium supplemented with 0.2% penicillin/streptomycin.
All cells were cultured at 37 °C in a 5% CO2 humidified atmosphere.
QM-7 cell culture: QM-7 cell lines of avian myogenic origin were cultured in high-glucose M199 medium (Gibco, USA) with 10% fetal bovine serum (FBS), 10% tryptose phosphate broth solution (Sigma, USA), and 0.2% penicillin/streptomycin.
CPM isolation and culture: Primary myoblasts were isolated from the leg muscle of 11-day old chicken embryos. First, the muscle tissues were dissected away from the skin and bone, and then homogenized in a petri dish. To release single cells, the suspension was digested with pancreatin for 20 min at 37 °C. After neutralization with complete medium, single cells were collected by centrifugation at 500 × g. Subsequently, serial plating was performed to enrich for primary myoblasts and eliminate fibroblasts. Primary myoblasts were cultured in Roswell Park Memorial Institute (RPMI)-1640 medium (Gibco, USA) with 20% FBS, 1% nonessential amino acids, and 0.2% penicillin/streptomycin. The differentiation of myoblasts was induced by RPMI-1640 medium supplemented with 0.2% penicillin/streptomycin.
All cells were cultured at 37 °C in a 5% CO2 humidified atmosphere.
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