Figure 6.

Number of Surviving Transplanted Cells Increased in Animals Pre-treated with Cyclosporin

(A) Timeline of experimental setup for hOPC transplantation in cyclosporin-treated versus cyclosporin-untreated grafted animals. (B) Representative images of the whole-mount surface preparation of cochlear epithelia at 14 days post-implantation. Dil-labeled cells (red) correspond to grafted hOPCs and nuclei were stained with DAPI (blue). (C) The number of Dil-labeled cells were quantified in cochlear sensory epithelium area at different cochlear turns. The CSE was represented in the space between dotted line in (B). Cyclosporin-treated group is represented in green color, and cyclosporin-untreated group is represented in orange color. Each floating bar represents group means (line) and min to max (box) values (n = 3). ANOVA test was used for multiple comparisons; different letters indicate significant differences between groups (**p < 0.005). (D) Virtual transversal images of cochlear sensory epithelium were obtained by 3D reconstructions of Z-stack series. Schematic reference to basilar membrane (BM) and reticular lamina (RL) is shown with dotted lines. The distance of Dil-labeled cells from basilar membrane was determined by Zen software. (E) The percentages of cells found at different distances was determined by the ratio of number of cells quantified in a delimited distance/total number of cells quantified into the entire turn of the cochlea. Data shown correspond to the basal turn of the cochlea from cyclosporin-treated (shown in green) and cyclosporin-untreated (shown in orange) animals. CSE, cochlear sensory epithelium; OHC, outer hair cell; IHC, inner hair cell; BM, basilar membrane; RL, reticular lamina. Scale bar, 20 μm.