GSM8587192: ApcMin ctrl biol rep 2, scRNAseq; Mus musculus; RNA-Seq
Macroscopic ApcMin/+ adenomas from all different anatomical regions of the small intestine were dissected and stored in high-glucose DMEM (Gibco, #11960-044) medium on ice. After isolation of all adenomas, the medium was replaced with a cell dissociation buffer containing 4 mg/ml dispase II (Sigma, #04942078001), 1 mg/ml collagenase I (Worthington, #LS004196), 1 mg/ml collagenase H (Roche, #11074032001), and 1000 U/ml benzonase (ChemCruz, #sc-202391) in HBSS (Gibco, #14175-053). After 30 min of incubation at 37°C, trypsin-EDTA (Thermo Fisher Scientific, #25200056) was added, and incubation was continued for 10 min at 37°C. The samples were vortexed regularly during incubation. Trypsin was inactivated in high-glucose DMEM. Samples were filtered through a 40 mm filter, followed by centrifugation for 10 min at 150 RCF. 10 000 cells in 0.04% BSA in HBSS with good viability (> 70%) per sample were targeted for analysis with Chromium X. Library was performed according to the manufacturer`s instructions (single cell 3' protocol, 10x Genomics)