Medulloblastoma Cell Culture Protocol

Patient derived human medulloblastoma lines D425 (Sigma-Aldrich, SCC290) as well as D283 and DAOY (ATCC, HTB-185 and HTB-186) were cultured in DMEM media supplemented with 10% FBS and 1% PEST (Sigma-Aldrich, P0781). Cells were cultured as monolayers on ultra-low attachment flasks and maintained at 37 °C in an incubation chamber with an atmosphere of 5% CO₂. These lines were chosen for subsequent cell proliferation and dose response assays and as human cell counterparts to our animal model cell lines due to their similarity – either in expressing MYC/MYCN or ARF status – transcriptional data regarding gene expression can be found within this paper and in our previously published work. Cell lines used were contamination-free when obtained from repositories or vendors but tested regularly upon culturing and passaging using a Mycoplasma detection kit, MycoAlert (Lonza). Only confirmed mycoplasma-free lines were used in research.