Establishing Glioblastoma Cell Lines from Patient Samples

320- and 1206-GSC cell lines were derived as previously described [15 (link)–17 (link)] and reused here. Briefly, after informed consent was obtained, tumor samples classified as glioblastoma, based on the World Health Organization (WHO) criteria, were collected from patients undergoing surgical treatment at the National Institutes of Health (NIH) or from Weill Cornell Medicine/New York Presbyterian Hospital in accordance with the appropriate Institutional Review Boards. Within 1–3 h after surgical removal, tumors were washed in PBS and enzymatically dissociated into single cells. Tumor cells were cultured in NBE medium consisting of neurobasal medium (Thermo Fisher Scientific), N2 and B27 supplements (Thermo Fisher Scientific), and human recombinant bFGF and EGF (25 ng/mL each; R&D Systems) plus Heparin sodium and L-Glutamine. Regular mycoplasma screening was performed using the MycoAlert Detection Kit (Lonza Inc.).