GSM5464743: Differentiation day 25; Mus musculus; RNA-Seq
Organoids from passage 2 (day 8) in expansion medium, from differentiated day 5, and from differentiated day 25 were collected and incubating with 0.125% trypsin for 25 min and dissociated into single cells by mechanical pipetting. Nuclei were stained with DAPI (Sigma) on ice for 10 minutes, then cells were sorted using a MoFlo® SX FACS cytometer (Beckman Coulter). Sorted single, living cells were resuspended in 40 mL of fresh cell media. Single-cell transcriptomic profiling was performed using the Single Cell 3' Reagent v3 kits according to the manufacturer's recommendations (10X Genomics