Microbead injection into the anterior chamber of C57BL/6 mice to induce elevation of IOP and glaucoma model.

<p>(A) After 4 weeks, microbeads can be detected in the Schlemm’s canal (orange fluorescence on the right side of the photograph). (B) Fitted geometric mean IOP changes with the 95% confidence limits in all microbead injected and control groups. (group 1–5). Saline-injected group showed mean IOPs of ≤ 10 mmHg (group 1). Microbead injected eyes (group 2–5) showed rapid and steady rise in IOPs peaking between 5–12 days and sustained elevated IOPs during the one-month study period. Data were analyzed by fitting a mixed effects model to the logarithm of the data with two fixed effects (treatment and time) and the subject as a random intercept. Multiple comparisons were conducted using the Tukey test on the fitted data. Asterisks denote statistically significant differences <i>versus</i> all groups within the same time point. (C) Representative retinal wholemount after intravitreal hNP transplantation imaged on confocal microscopy. Red fluorescent structures (ß-III tubulin) represent host RGCs and their nerve fibers. (D) Green fluorescent dots (FITC, HLA Class I antigen expression) present hNPs that have penetrated the host retina. (E) High-resolution images of retinal flatmounts. (F, G) hNPs expressing IGF-TD can also be detected by their expression of IGF-1 (FITC-green) and TD (Rhodamine-red) components of the fusion protein. (H) Merged image of F and G. Abbreviations: ONL, outer nuclear layer; INL, inner nuclear layer; RGCs, retinal ganglion cells. IGF-TD, transplanted hNP<sup>IGF-TD</sup> cells after microbead injection; TD, transplanted hNP<sup>TD</sup> cells after microbead injection. hNP, untransfected hNPs after microbead injection; SA, intravitreal saline (no cells) injection after microbead injection; NO, intravitreal saline injection and saline injection into the anterior chamber (no microbead and cell injection).</p>