Fig. 2

Influence of immunosuppressant drug on the long-term metabolic activity of retinal organoids. 2P microscopy (fluorescence lifetime [FLIM] analysis) was used to show shift from glycolytic to OXPHOS based metabolism, measured as ratio of free to bound NADH. a and b Two different sets of organoids were evaluated after 1-week and 4-week exposure to MPA (red, 0.5 μg/ml), TAC (green, 3 ng/ml) or a combination of both (purple) at the lowest effective concentration reported for human clinical trials. No significant difference in free/bound NADH was detected (one-way ANOVA) between drug-treated organoids and controls. c Overview of average free/bound NADH profiles among RO lots across days 50–170 of differentiation. Grey data points represent a modified format from Fig. 1b of previous publication [5], control groups for drug treatment are colored in blue. Control samples appear to correspond to the previously reported samples in [5]. d mRNA expression heatmap across IS drug exposed ROs. TAC and MPA treated samples showed similar gene expression level as control. The combined treated sample showed a discernable fold change in downregulation of CRX and NRL compared to the other three groups. Retina organoids’ age ranged from 13.5 to 16.5 weeks (day 95–115). Drug-treated for 4 wks. log2 F.E − log2(fold expression). Ctrl = control; MPA = Mycophenolic acid (0.5 µg/ml); TAC = Tacrolimus (3 ng/ml)