Fig. 2

Transplantation of human photoreceptors to investigate material transfer. A Retinal organoids (derived from the nuclear CRX-GFP H9 reporter cell line) were dissociated at week 10, 14, and 20, FAC-sorted to isolate CRX-GFP⁺ photoreceptors and transplanted into adult C57BL/6J, Nrl^−/− or NSG mice. Tissue was collected 21 days later to look for evidence of material transfer. B Representative section of a week 8 retinal organoid with detectable GFP reporter (green) co-localized with CRX (red). Scale bar = 100 μm. C Quantification of CRX-GFP photoreceptor population in retinal organoids dissociated and sorted at weeks 10, 14, and 20. Live photoreceptors were gated for GFP (FITC) on the x-axis with an empty channel on the y-axis (BV421) to gate out weak auto-fluorescent cells. Data presented as mean ± SEM, analyzed by one way ANOVA with Tukey’s post-hoc. *p < 0.05; **p < 0.01; ****p < 0.0001 Sorts were performed on the Aria IIIu, BD Influx, and Aria III. D Criteria for donor cell identity and material transfer. E Donor human photoreceptors are distinguished by their significantly larger nuclei. Data presented as mean ± SEM (n = 57–114 nuclei from at least 3 animals in each group), one-way ANOVA with Tukey’s post-hoc. ****p < 0.0001