Dual tumour–myeloid targeting of glioblastoma with GPNMB CAR-T cells

Cell lines and patient-derived GSC cultures Human GBM samples were obtained from patients who provided written informed consent for tissue collection and use in research and publication, under the approval of the Hamilton Health Sciences and McMaster Health Sciences Research Ethics Board (no. 16078). Tumour samples were processed using established protocols 60 . GBM cells were cultured in Neurocult Complete (NCC) medium, a commercially available serum-free NSC medium (STEMCELL Technologies, 05751), supplemented with human recombinant epidermal growth factor (20 ng ml −1 ; STEMCELL Technologies, 78006), basic fibroblast growth factor (20 ng ml −1 ; STEMCELL Technologies, 78006), heparin (2 μg ml −1 ; STEMCELL Technologies, 07980) and antibiotic–antimycotic solution (1×; Wisent, 450-115-EL). NSCs were cultured and maintained using similar protocols as described previously 61 . SK-MEL-2 cells, MDA-MB-231 and HEK293T cells were purchased from the American Type Culture Collection (ATCC) and grown in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% FBS and 1% non-essential amino acids (NEAA; Thermo Fisher, 11140050). NHAs were purchased from ATCC and grown in DMEM/F12 medium (Gibco, 11320033) supplemented with 10% FBS, and epidermal growth factor and fibroblast growth factor as above. …