Neonatal human fibroblasts (HFF-1; ATCC), 293T and the human glioma lines U-87 MG and LN229 (ATCC) were cultured in DMEM containing 10% FBS, 2 mM GlutaMAX (Invitrogen), 50 U ml−1 penicillin and 50 mg ml−1 streptomycin (Invitrogen). Human foreskin fibroblasts (HFF) were grown in collagen I coated plates (BD biosciences). Human iPSCs were cultured in chemically defined hES/hiPSCs growth media, mTeSR on growth factor reduced matrigel (BD biosciences) coated plates. Briefly, 70–80% confluent iPSCs cells were treated with dispase (Invitrogen) for 7 min at 37 °C and the colonies were dispersed to small clusters and lifted carefully using a 5-ml glass pipette at a ratio of ∼1:4. GTICs, previously reported and characterized5, were obtained from BioRep S.r.l. (NS27Z +B AB0789) and maintained in culture as described elsewhere5. All cell lines were maintained in an incubator (37 °C, 5% CO2) with media changes every day (iPSCs) or every second day (fibroblasts, 293T cells, GTICs).
