Illumina HiSeq 2500 paired end sequencing; Gene expression profiling of core glioma stem cells: a novel glioblastoma cell population with diminished antigen presentation
Gene expression profiling of core glioma stem cells: a novel glioblastoma cell population with diminished antigen presentation
ic-GSC were splited with ReLeSR™, a passaging reagent ic-GSC were cultured on coated dishes with Matrigel containing the supplemented mTeSR1 media with daily media change Lentivirus TetO-FUW-OSKM and FUW-M2rtTA were generated in 293T cells, in mTeSR1 media. The viral supernatants were mixed at 1:1 ratio and filtered. GSCs were seeded at a density of 0.3 x 106 per well of a 6-well plate, and infected four times at 8h, 24h, 32h, and 48h, with 2 μg/ml. GBM-DCs were treated with doxycycline 2g/ml to induce OSKM expression. ic-GSC colonies appeared after four weeks and single-cell clone selection were manually generated in 96-well plate. We selected three ic-GSC clones. RNA was obtained using the EZNA Total RNA Kit I Samples were processed using the TruSeq RNA Sample Prep v3 kit (Illumina) according to the manufacturers instructions.