Xenograft tumors were dissociated using papain (Worthington Biochemical Corporation, Lakewood, NJ) and cultured overnight in supplemented neurobasal medium (neurobasal medium (Life Technologies) with 2% B27 (Life Technologies), 1% penicillin/streptomycin (Life Technologies), 1 mM sodium pyruvate (Life Technologies), 2 mM L-glutamine, 20 ng/mL EGF (R&D Systems, Minneapolis, MN, USA), and 20 ng/mL FGF-2 (R&D Systems)). T4121, T3691, and T387 xenografts were sorted for CD133+ (CSC) and CD133-(non-CSC) populations using the CD133 Magnetic Bead Kit for Hematopoietic Cells (CD133/2; Miltenyi Biotech, San Diego, CA, USA). CD133+ cells were maintained in supplemented neurobasal. CD133- cells were maintained in DMEM with 10% FBS and 1% pen/strep. L2 cells were maintained in these divergent media conditions without sorting.
HeLa and NIH 3T3 cells were maintained in DMEM with 10% FBS and 1% pen/strep. The HeLa-Cx46 stable cell line was cultured with the addition of 400 mg/mL G418. All cells were grown in a humidified incubator at 37 C with 5% CO2.
HeLa and NIH 3T3 cells were maintained in DMEM with 10% FBS and 1% pen/strep. The HeLa-Cx46 stable cell line was cultured with the addition of 400 mg/mL G418. All cells were grown in a humidified incubator at 37 C with 5% CO2.