title: GSM8190282: hPD organoids 1, live sort, rep3, scRNAseq; Homo sapiens; RNA-Seq
library_construction_protocol: The islet-depleted tissue was mechanically dispersed using a 21G needle (BD Microlance 3), followed by three washes with Hank's Balanced Salt Solution (Thermo Fisher Scientific) supplemented with 100μg/ml trypsin inhibitor (Merck) at 200g for 4 minutes at 4°C. The tissue was dissociated in TrypLE at 37°C 15-30 minutes depending on the sample. Dissociation was stopped with Advanced DMEM/F12 (Thermo Fisher Scientific) supplemented with 10% Fetal Bovine Serum (FBS; Greiner Bio-One). The cells were centrifuged at 200g for 3 minutes, at 4°C to collect the cell pellet and resuspended in DPBS (no calcium, no magnesium; Thermo Fisher Scientific) supplemented with 2% FBS and 1mM EDTA (Thermo Fisher Scientific) and passed through a 40μm cell-strainer before FACS sorting into 384 well cell capture plates. hPDO's were collected and dissociated for 15 minutes in TrypLE, washed in DPBS, strained and sorted into 384 cell capture plates. CEL-seq2 (Hashimshony et al 2016)