Cell incubator

HEK293T cells: HEK293T cells were cultured in DMEM High-Glucose medium (Sigma-Aldrich, St. Louis, MO, USA) containing 10% fetal bovine serum (Gibco, Grand Island, NY, USA) and 1% penicillin–streptomycin–amphotericin (Solarbio, Beijing, China).
DF-1 cells: DF-1 cells were cultured in DME-F12 (Sigma-Aldrich, St. Louis, MO, USA) medium containing 10% fetal bovine serum and 1% penicillin–streptomycin–amphotericin.
Isolation and culture of chicken primary myoblasts: The leg muscles of 11/12-day-old chick embryos were taken. The bones and blood clots in the muscles were separated, and the muscles were evenly cut into a 10 cm cell culture dish. The muscle was digested with collagenase type I (Gibco, Grand Island, NY, USA), at 37 °C, to obtain single cells. The cell suspension after digestion was filtered and centrifuged, and then cultured in a 10 cm cell culture dish. Fibroblasts were removed after three times differential attachment. The growth medium was DME-F12, containing 20% fetal bovine serum and 1% penicillin–streptomycin–amphotericin. The differentiation medium was DME-F12 containing 4% fetal bovine serum and 1% penicillin–streptomycin–amphotericin. All cells were cultured in a cell incubator (Binder, Tuttlingen, Germany), with 5% CO₂, at 37 °C.

The human primary dermal fibroblast cell lines, GMO5565 (3-year-old male), GMO1651 (13-year-old female) and GMO5567A (12-year-old male) were all obtained from the Coriell Institute for Medical Research (Camden, NJ, USA). HGPS cell lines were obtained from the Progeria Research Foundation Cell and Tissue Bank. The following HGPS cell lines were used: HGADFN003 (2-year-old male) and HGADFN127 (3-year-old female). The 3T3-L1 (ATCC^® CL-173™) cell line was purchased from ATCC (Manassas, VA, USA). Fibroblast cell lines were cultured as monocultures in DMEM (Thermo Fisher—Gibco, Waltham, MA, USA, D6429) supplemented with 15% fetal bovine serum (FBS) (Thermo Fisher—Gibco, 10270106), 1% L-glutamine (Thermo Fisher—Gibco, 25030081), 1% penicillin/streptomycin (Thermo Fisher—Gibco, 1514022) and 0.5% gentamycin (Thermo Fisher—Gibco, 15710049). The fibroblasts were subcultured and used when they were approximately 80% confluent. All cultures were performed in a cell incubator (Binder, Tuttlingen, Germany, 9140-0046) in a humidified chamber at 37 °C and 5% CO₂.