Figure 2.

Direct transfection of organoids in Matrigel microbeads. (A) Experimental protocol for direct transfection of encapsulated RWPE-1-GFP organoids. Encapsulated RWPE-1-GFP cells were allowed to develop into organoids for 4 days and were then transfected with siRNA (control siAllStars and siGFP, 20 nM) for 3 days before analysis. (B) After 3 days of transfection by comparing different techniques, cells were extracted from microbeads and analyzed via flow cytometry. GFP knock-down was determined using the GFP MFI measurement as follows: GFP knock-down = 100 − (MFI siGFP/MFI siAllStars) × 100. Results are expressed as the percentage of GFP signal extinction and represent the mean value ± SEM of three experiments (*P < 0.05). (C) Cell viability was measured via trypan blue dye exclusion staining.