Medulloblastoma Cell Line Establishment and Culture

The human ONS-76 medulloblastoma cell line was obtained from the RIKEN Cell Bank (Tsukuba, Ibaraki, Japan). ONS-F8 and ONS-B11 cell lines were established from ONS-76 after irradiation as previously described [26 (link)]. The cells were cultured in minimal essential medium (MEM; Sigma-Aldrich Inc., Tokyo, Japan) containing 10% fetal bovine serum (FBS; Nichirei Biosciences Inc., Tokyo, Japan), 100 mg/ml streptomycin, and 100 U/ml penicillin (Sigma-Aldrich). Cells were incubated in a humidified atmosphere at 37°C with a 5% CO₂ atmosphere. For subcultures, cells were rinsed with Ca²⁺- and Mg²⁺-free PBS (Sigma-Aldrich), and dispersed with 0.25% trypsin containing 0.5 mM ethylenediaminetetraacetate (EDTA; Sigma-Aldrich). A final concentration of 50 mM DCA (Sigma-Aldrich) was added 48 h before analysis. The number of cells was determined with TC10^™ (Bio-Rad, Tokyo, Japan).