Abstract
Abstract: :
Purpose: To place ciliary body–derived, progenitor cells into the glaucoma–damaged retina of experimental rats and to direct their development into mature retinal neurons. Methods: The ciliary body epithelial cells of FVB.Cg–Tg(GFPU)5Nagy mice transgenic for Green Fluorescent Protein driven by the beta actin promoter were dissected immediately after sacrifice, dissociated by trypsin digestion, and allowed to multiply in EGF/FGF2 medium for 7 days (Tropepe et al, 2000). Proliferated spheres of daughter cells that developed were dissociated, and 15,000 cells were injected into each vitreous cavity of 17 Wistar rat eyes that had: 1) unilateral experimental glaucoma alone, 2) unilateral glaucoma plus retinal needle puncture, 3) retinal needle puncture alone, or 4) no retinal injury. After 4 weeks, retinas were harvested and studied in whole mount and cross–section by fluorescence microscopy. Results: Incorporation of GFP–labeled mouse progenitor cells was 1.6—3.7 times greater in glaucoma retinas than in non–glaucoma eyes (26.1 +/– 6.3 vs. 7.0 +/– 2.1, mean/SE, n = 6 & 7, p < 0.01, superficial retina), but (contrary to previous reports) lower by 50% in punctured retinas compared to non–punctured ones (18.8 +/– 10.5 vs. 45.6 +/– 6.1, n = 5 & 8, p < 0.04, ganglion cell layer). The density of progenitor incorporation was more than twice as high in the retina near the horizontal meridian than in the retina periphery (44.5 +/– 11.2 vs. 19.9 +/– 4.8, n = 13, p = 0.05). The number of progenitor cells incorporated into inner retinal layers was estimated as 7% of the number injected intravitreally. Conclusions:Progenitor cells resident in the adult rodent ciliary body multiply in culture and incorporate in significant numbers into rodent eyes with experimental glaucoma damage.
Keywords: regeneration • ganglion cells • pathology: experimental