R28 cell transplant in the RCS retina

Abstract

Abstract: : Purpose: To locate Di–I labeled retinal precursor cell line R–28 cells transplanted into the posterior retinas of dystrophic Royal College of Surgeons rats (dRCS). Methods: 4 week old dRCS were anesthetized with ether and injected with 10µl of 5x104 cells/µl R–28 cell suspension in PBS. The injection was into the subretinal debris zone of the superior temporal quadrant of the right eye of the rat using a 25 gauge sterilized needle with a wire insulation collar and a 1 ml syringe. Rats were allowed to recover and were sacrificed 4 weeks later by anesthetization with ether followed by exsanguination by perfusion with PBS and then 10% formalin in 0.1M sodium phosphate buffer, pH 7.4. Eyes were harvested and frozen sections of the entire globe were made in a cryostat. Slides were examined with a fluorescent microscope and digital images were taken. Results: No infection or cell rejection symptoms were noted. R28 cells settled into different regions of the eye. In separate animals, the cells aligned along the pigmented epithelium, the inner border of the retina, the outer border of the lens, and within the lens. Conclusions: R28 cells were able to survive and migrate within the dystrophic RCS retina. Further studies are needed to determine if differentiation of the cells occurred.

Keywords: transplantation • retinal degenerations: hereditary • microscopy: light/fluorescence/immunohistochemistry