Abstract
To develop biological approaches to restore vision, we developed a method of transplanting stem cell-derived retinal tissue into the subretinal space of a large-eye animal model (cat). Human embryonic stem cells (hESC) were differentiated to retinal organoids in a dish. hESC-derived retinal tissue was introduced into the subretinal space of wild-type cats following a pars plana vitrectomy. The cats were systemically immunosuppressed with either prednisolone or prednisolone plus cyclosporine A. The eyes were examined by fundoscopy and spectral-domain optical coherence tomography imaging for adverse effects due to the presence of the subretinal grafts. Immunohistochemistry was done with antibodies to retinal and human markers to delineate graft survival, differentiation, and integration into cat retina. We successfully delivered hESC-derived retinal tissue into the subretinal space of the cat eye. We observed strong infiltration of immune cells in the graft and surrounding tissue in the cats treated with prednisolone. In contrast, we showed better survival and low immune response to the graft in cats treated with prednisolone plus cyclosporine A. Immunohistochemistry with antibodies (STEM121, CALB2, DCX, and SMI-312) revealed large number of graft-derived fibers connecting the graft and the host. We also show presence of human-specific synaptophysin puncta in the cat retina. This work demonstrates feasibility of engrafting hESC-derived retinal tissue into the subretinal space of large-eye animal models. Transplanting retinal tissue in degenerating cat retina will enable rapid development of preclinical in vivo work focused on vision restoration.
Highlights
Vision is by far the most dominant and most important sense to all primates, including humans, with 80% or more of all sensory information being perceived by means of sight [1,2,3,4]
The retinal pigment epithelial layer was detected by immunolabeling with tight junction protein zonula occludens (ZO)-1 [103] and pigmented RPE marker PMEL17 [70,104] (Fig. 1g–n and Supplementary Fig. S1)
Photoreceptor progenitors were present on the apical side of the Human embryonic stem cells (hESC)-derived retinal tissue, whereas amacrine cells were restricted to the basal side
Summary
Vision is by far the most dominant and most important sense to all primates, including humans, with 80% or more of all sensory information being perceived by means of sight [1,2,3,4]. New and effective treatments to restore and preserve vision in patients with retinal degeneration (RD) are urgently needed. Similar to patches of retina preserved by gene augmentation therapy, the islands of transplanted mutation-free retina have the potential to survive, and with synaptic connectivity to the host, at least partially restore visual function [15,16,18,19,37,38,39,40]. The progressive nature of RD conditions, where the dying PRs destroy RD retinal matrix and trigger the death of healthy PR around them [41,42,43,44,45], indicates that transplanting dissociated mutation-free retinal cells into the degenerating
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