Culturing Mouse Motor Neurons and Cell Lines

HyClone™ Dulbecco’s modified Eagle’s medium (DMEM) (GE Healthcare Life Science) with 10% fetal bovine serum (FBS) (Corning) was used for mouse motor neuron-like hybrid cell line (NSC-34), PC12 cells, HEK293T, and BMSCs culture. The cell lines of NSC-34, PC12, and HEK293T were bought from American Type Culture Collection (ATCC, USA). PC12 cells were incubated by culture medium containing 50 ng/ml NGF (Peprotech, 450-34) for 24 h and keep passage for the following experiment. BMSCs were isolated from the fetuses of SD rats as previously described [21 (link)]. All cells were maintained at 37 °C in a humidified incubator with 5% CO₂ with cell passage performed, when cell density was ~ 90%. Cryopreservation solutions used comprised of complete medium and 10% dimethyl sulfoxide (DMSO).