GSM9391952: sample 3 - TECs from Responder  tumours; Mus musculus; RNA-Seq
Tumours were removed and cut to small pieces with a scissor. Then they were digested using 0.1% collagenase type I diluted in DMEM (Gibco), supplemented with 10% of EGM2 media (Promocell), 1% of BSA (Sigma), DNAse I, 100 mM HEPES and 2 mM EDTA. First the mix of tumour pieces was mixed with the digestion media, and tumours were processed in gentleMACS™ Dissociator (Miltenyi), then incubated at 37 degrees for 30 min, and then processed again in the Dissociator. Homogenized tumours were filtered with a 70-um filter and then the cell pellet was subjected to different procedures. Endothelial cells were isolated using cell death removal and EC isolation bead kits from Miltenyi. To obtain a pure population of ECs, we selected DAPI-, CD45-, EPCAM-, CD31+ cells by FACs. Cells were immediately processed for scRNAseq. Single-cell suspensions were prepared and Live CD45- CD31+ populations were sorted. After this, cells were loaded and processed on the 10X Chromium Controller using the CG000331 Chromium Next GEM Single Cell 5' Reagent Kits v2 (Dual Index) User guide enabled us to process the gene expression and immune profiling of the samples. Each library can be recognized by its unique sample index. Sample sequencing was performed at Novogene using a Novaseq 6000 platform and PE150 sequencing strategy.
GEO
GSM9391952_r1