GSM9437895: Porcine intestinal organoid 3 day; Sus scrofa; RNA-Seq
After organoid cells culture for 1, 3, and 5 days, treated with TrypLE (Thermo Fisher Scientific, Wilmington, DE, USA) containing 10 μM Y-27632 for 20 min, with pipetting every 5 min to obtain single-cell form. Dead cells were removed from a Dead cell removal kit (Miltenyl Biotec, Bergisch Gladbach, Germany). Single cells were washed in PBS containing 0.04% BSA and counted using a Countness II (Thermo Fisher, Waltham, MA, USA) to determine their cell number. To obtain single cell RNA-sequencing libraries, Chromium Next GEM Single Cell 3' reagent kit v3.1 (10X Genomics, Pleasanton, CA, USA) was used according to the manufacturer's protocol. the cells were diluted into the Chromium Next GEM Chip G as to yield a recovery of ~ 5,000 single-cell and then the cell libraries were sequenced using a multiplex on the Novaseq 6000 sequencer (Illumina, San Diego, CA) to produce on average a minimum of 60,000 reads per single cell.