Cell Culture Protocols for Research

HeLa, SH-SY5Y, 293T, and U2OS cells were all purchased from the National Collection of Authenticated Cell Cultures (Shanghai, China). They were cultured in DMEM-high glucose (Gibco, Carlsbad, CA, USA), which was supplemented with 10% (v/v) fetal bovine serum (FBS, Gibco, Carlsbad, CA, USA) and 1% penicillin/streptomycin at 37 °C in 5% CO₂. In addition, human embryonic stem cells (ESCs) H9 were obtained from Dr. Hongkui Deng, Peking University Stem Cell Research Center (Beijing, China) [17 (link)]. Coat plates with 1% matrigel (354277, Corning, New York, NY, USA) were heated at 37 °C for 1 h, cultured in mTeSR™1 medium (85850, STEMCELL, Vancouver, Canada), and supplemented with 1% penicillin/streptomycin and Y27632 (10 μM).