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. 2020 Oct 20;28(3):1041–1061. doi: 10.1038/s41418-020-00636-4

Fig. 2. Exosomes mediates therapeutic effects of mesenchymal stem cell transplantation (MSCT) on N-methyl-N-nitrosourea (MNU)-induced photoreceptor apoptosis and retinal degeneration.

Fig. 2

Representative hematoxylin and eosin (H&E) staining images of retinal tissues (A) and the corresponding quantitative analysis of outer nuclear layer (ONL) thickness (B). Mice were injected with MNU with or without MSCT. GW4869, a neutral sphingomyelinase inhibitor for blocking exosome generation, which was used for MSC preconditioning before transplantation; GCL ganglion cell layer, INL inner nuclear layer, ONH optic nerve head. Scale bars = 50 μm. *P < 0.05 by the Kruskal–Wallis test for area under curve (AUC). N = 3 per group. Representative scotopic electroretinography (ERG) waveforms (C) and the corresponding quantitative analyses of amplitude changes of a-wave (D) and b-wave (E). *P < 0.05 by the Kruskal–Wallis tests. N = 6 per group. Representative photopic ERG waveforms (F) and the corresponding quantitative analysis of b-wave amplitude changes (G). *P < 0.05 by the Kruskal–Wallis tests. N = 6 per group. Representative H&E staining images of retinal tissues (H) and the corresponding quantitative analysis of ONL thickness (I). Ctrl control, no MNU treatment, EXOT exosomal transplantation after MNU injection. Scale bars = 50 μm. *P < 0.05 by the Kruskal–Wallis test for AUC. N = 3 per group. Representative scotopic ERG waveforms (J) and the corresponding quantitative analyses of amplitude changes of a-wave (K) and b-wave (L). *P < 0.05 by the Kruskal–Wallis tests. N = 6 per group. Representative photopic ERG waveforms (M) and the corresponding quantitative analysis of b-wave amplitude changes (N). *P < 0.05 by the Kruskal–Wallis tests. N = 6 per group. Representative terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL, green) staining images of retinal tissues counterstained by Hoechst 33342 (blue) (O) and the corresponding quantitative analysis of percentages of TUNEL+ cells over total ONL cells (P). Scale bars = 50 μm. *P < 0.05 by the Kruskal–Wallis test. N = 3 per group. Representative flow cytometric images showing death of 661W cone photoreceptor cells (Q) and the corresponding quantitative analysis of percentages of apoptotic (Annexin V+PIplus Annexin V+PI+) 661W cells (R). The MSC group, Transwell co-culture of MNU-treated 661W cells with MSCs; the GW4869 group, GW4869 was used for MSC preconditioning before co-culture; the EXO group, MSC-derived exosomes were added to MNU-treated 661W cells. PI propidium iodide. *P < 0.05 by the Kruskal–Wallis test. N = 3 per group. Data represent median ± range for (B), (I), (P), and (R). Data are represented as box (25th, 50th, and 75th percentiles) and whisker (range) plots for (D), (E), (G), (K), (L), and (N).