GSM7765122: Day 5 HyA-FMB organoid, donor 1, scRNAseq; Homo sapiens; RNA-Seq
Control and HyA-FMB organoids from days 1-10 and ectopic HyA-FMB transplant were digested in high-glucose DMEM containing 0.2% collagenase II, 0.1% hyaluronidase, and 0.1% dispase with gentle pipetting every 20-30 minutes.  Enzymatic digestion was observed in 6 well-plates, and digestion was terminated when aggregates were completely dissolved, for up to 2 hours.  Enzyme was inhibited with 20% FBS, and cells were filtered through 70 µm strainers and re-suspended in PBS supplemented with 0.04% high-quality bovine serum albumin at 1000 cells/µL for single-cell capture.  Naïve hBMSC/SSC controls (from donor 3) required a 10-minute incubation with collagenase II on monolayer cultures prior to cell detachment and cell capture. Single-cell RNA sequencing was performed using 10X Genomics Chromium instruments.