Automating iPSC generation to enable autologous photoreceptor cell replacement therapy

scRNAseq profiling of retinal organoids derived from iPSCs generated on a robot platform. Approximately ten representative organoids were selected from culture. Organoids were allowed to settle by gravity in a 1.5 mL tube and medium was aspirated. Organoids were then dissociated with 20 units/mL of papain (Worthington Biochemical Corporation, Lakewood, NJ) with 60 units/mL DNase I (Worthington Biochemical Corporation) on a shaker at 37˚C for approximately 60 minutes. Following dissociation, cells were pelleted at 400 x g for 5 minutes, resuspended in dPBS -/- (Thermo Fisher Scientific) with 0.04% non-acetylated bovine serum albumin (New England Biolabs, Ipswich, MA), and processed immediately for encapsulation and barcoding using the Chromium Controller (10X Genomics, Pleasanton, CA).