GSM5026213: organoid_d0_scRNAseq_2; Homo sapiens; RNA-Seq
Organoids were rinsed with DPBS, collected into 1.5 ml conical tube containing 500 ml TrypLE Select and incubated for 5 min at 37°C, passed 3 to 4 times through a 27G needle, and incubated for 5 min at 37°C after adding additional 500 ml of TrypLE Select. Organoids were dissociated into single cell suspensions by passing 3 to 4 times through a 30G needle, followed by centrifugation at 600xg for 5 min. Supernatants were discarded, and cell pellets were resuspended in DPBS and filtered through a 40 mm cell strainer. Cell numbers were quantified on a Cellometer cell counter and adjusted to 1000 cells/ml in DPBS. Single cell cDNA library preparation was performed following manufacturer's protocols (https://www.10xgenomics.com). Single cell suspension were loaded on a Chromium controller to generate single cell GEMS (Gel Beads-In-Emulsions). The scRNA-Seq library was prepared with chromium single cell 3' reagent kit v3 (10x Genomics)