title: RNA-seq of 4 naive GBM cell lines upon dexamethasone (0.1 uM) treatment
design: RNA-seq of 4 naive GBM cell lines upon dexamethasone (0.1 uM) treatment
library_name: 5B_p
library_construction_protocol: GBM patient-derived cell lines were obtained as described in \"Oudin, A. et al. Protocol for derivation of organoids and patient-derived orthotopic xenografts from glioma patient tumors. STAR Protocols 2, 100534, doi:10.1016/j.xpro.2021.100534 (2021).\". Intracortical implantation of organoids in immunodeficient mice is performed to induce tumor growth in vivo and to establish PDOX models method was used. Animals must be housed in individually ventilated cages in a Specific Pathogen Free (SPF) facility, under controlled environment (temperature 22 ± 2°C, humidity between 45% and 65% and 12 h light / 12 h dark cycle) with free access to autoclaved and acidified water and irradiated food ad libitum. GBM naive cells were seeded at different densities in 6-well plates and cultured in Plasmax. 24h after seeding, media were renewed with fresh Plasmax, supplemented with vehicle or dexamethasone 0.1 uM, and cells were incubated for 72 hours at 21% O2. Cells were then harvested for RNA extraction. Samples were processed using the QIAshredder/RNeasy Mini Kit (Qiagen) Libraries for cluster generation and DNA sequencing were prepared using Illumina TruSeq RNA Library Preparation Kit. Quality and quantity of the DNA libraries were assessed on an Agilent 2200 TapeStation (D1000 screentape) and Qubit (Thermo Fisher Scientific), respectively.