Differentiation of Human Embryonic Stem Cell–Derived Retinal Progenitors into Retinal Cells by Sonic Hedgehog and/or Retinal Pigmented Epithelium and Transplantation into the Subretinal Space of Sodium Iodate–Injected Rabbits

Abstract

Transplantation of retinal cells has recently provided a promising therapeutic approach for retinal degeneration. Here, we differentiated initially retinal progenitors (RPs) from adherent feeder-free human embryonic stem cells (hESCs) with the use of defined media supplemented with a specific combination of growth factors. The differentiated RPs highly (>80%) expressed related molecular features that included Six3 at an early stage in addition to Crx, Rx, Pax6, Otx2, and Chx10 at later stage. Next, we examined the induction of photoreceptors by Shh and/or the coculture of rabbit retinal pigmented epithelium with hESCs-derived RPs. The differentiation of retinal cells was demonstrated by protein and gene expression in all groups. However, S-Opsin, a cone photoreceptor marker, had higher expression in the presence of Shh, whereas expressions of Gli and Hes1 decreased in the same group. Finally, hESC-derived RPs were treated with Shh transplanted into the subretinal space of sodium iodate-injected albino-type adult rabbits and analyzed 4 weeks later. Transplanted retinal cells survived, migrated into retinal layers, and restored a small but significant B-wave. The grafted cells expressed photoreceptor markers, S-Opsin and Rhodopsin. Our results indicate that putative hESC-derived retinal cells express related genes and proteins. Further, our results show that retinal-like cells can be useful replacements for photoreceptors in retinal diseases.

Stem cell based therapies for age-related macular degeneration: The promises and the challenges

“…The authors reported that these ES-derived RPE-like cells expressed typical RPE markers, such as ZO-1, RPE65, CRALBP, and Mertk, had extensive apical microvilli, and were able to phagocytize latex beads. The cells enhanced photoreceptor survival upon transplantation into the subretinal space of Royal College of Surgeons (RCS) rats, a well-established model of AMD (Amirpour et al, 2012). Haruta et al (2004) had originally reported successful transplantation of ES-derived RPE cells into the RCS rat subretinal space with enhanced survival of host photoreceptor cells (Haruta et al, 2004).…”

Multimodal Fundus Imaging of Sodium Iodate-Treated Mice Informs RPE Susceptibility and Origins of Increased Fundus Autofluorescence

“…3–9,11 The work presented here has further advanced our understanding of this model. Using both in vitro and mouse models, interactions between the oxidative effects of NaIO 3 on RPE and photooxidative stress imposed by lipofuscin bisretinoid also have been revealed.…”

“…1–6 In recent years, NaIO 3 administration in rats and mice has been used to model the atrophic lesions that are a feature of AMD 7 and has been favored as a means to denude RPE in advance of cell therapy. 4,5,8–10 …”

Subretinal Injection: A Review on the Novel Route of Therapeutic Delivery for Vitreoretinal Diseases

“…In rats, photoreceptor degeneration can be prevented by subretinal transplantation of human fetal lung fibroblasts expressing the ciliary neurotrophic factor gene [66] , and laser-induced choroidal neovascularization can be inhibited by subretinal transplantation of RPE overexpressing fibulin-5 [67] . Of all of the cell types, progenitor cells and stem cells, such as human neural progenitor cells [68] , human retinal progenitor cells [69] , progenitor cells from the porcine neural retina [70] , forebrain progenitor cells [71] , brain-derived precursor cells [72] , human embryonic stem cell-derived retinal progenitors [73] , human RPE stem cells [74] , human bone marrow mesenchymal stem cells [75] , and rat mesenchymal stem cells [76] , are the most popular when given subretinally for cell replacement therapy for retinal degeneration. All these cells are considered to have the capability to survive and migrate into retinal layers and restore retina function or induce cell regeneration in different types of retinal cells when delivered via the subretinal route.…”