GSM9421940: a2_2024 CSPL; Homo sapiens; RNA-Seq
Research biopsies were immediately immersed in RPMI containing 1 mM DTT, 20 mM EDTA and 2% FBS at 37o C for 15 min to isolate epithelial cells and dissociated in Liberase TL (200 μg/ml of Liberase TL) containing solution in RPMI for 20 min at 37°C. Digested cell suspensions were passed through a 100-μm cell strainer at least three times, washed with complete media, and combined with the epithelial fraction before processing for droplet-based single-cell RNA-seq (scRNA-seq). To ensure 10,000 cells were encapsulated into droplets using 10x Chromium GEM technology single cells were suspended at 1,500 cells/ microliter concentration. Libraries were prepared in house using the Chromium Next GEM Single Cell 3′ Reagent Kits v.3.1and 5' reagent Kits v2 (10x Genomics). scRNA-seq libraries were sequenced using the Nova-Seq system (Illumina). 10x Genomics Chromium Next GEM single cell 3' reagent kits v3.1 (Dual index) and 10x Genomics Chromium Next GEM single cell 5' reagent kits v2 (Dual index).