Abstract
PurposeTo study if human embryonic stem cell–derived photoreceptors could survive and function without the support of retinal pigment epithelium (RPE) after transplantation into Royal College of Surgeons rats, a rat model of retinal degeneration caused by RPE dysfunction.MethodsCSC14 human embryonic stem cells were differentiated into primordial eye structures called retinal organoids. Retinal organoids were analyzed by quantitative PCR and immunofluorescence and compared with human fetal retina. Retinal organoid sheets (30–70 day of differentiation) were transplanted into immunodeficient RCS rats, aged 44 to 56 days. The development of transplant organoids in vivo in relation to the host was examined by optical coherence tomography. Visual function was assessed by optokinetic testing, electroretinogram, and superior colliculus electrophysiologic recording. Cryostat sections were analyzed for various retinal, synaptic, and donor markers.ResultsRetinal organoids showed similar gene expression to human fetal retina transplanted rats demonstrated significant improvement in visual function compared with RCS nonsurgery and sham surgery controls by ERGs at 2 months after surgery (but not later), optokinetic testing (up to 6 months after surgery) and electrophysiologic superior colliculus recordings (6–8 months after surgery). The transplanted organoids survived more than 7 months; developed photoreceptors with inner and outer segments, and other retinal cells; and were well-integrated within the host.ConclusionsThis study, to our knowledge, is the first to show that transplanted photoreceptors survive and function even with host's dysfunctional RPE. Our findings suggest that transplantation of organoid sheets from stem cells may be a promising approach/therapeutic for blinding diseases.
Highlights
Title Retina Organoid Transplants Develop Photoreceptors and Improve Visual Function in Royal College of Surgeons (RCS) Rats With retinal pigment epithelium (RPE) Dysfunction
Retinal organoids showed similar gene expression to human fetal retina transplanted rats demonstrated significant improvement in visual function compared with RCS nonsurgery and sham surgery controls by ERGs at 2 months after surgery, optokinetic testing and electrophysiologic superior colliculus recordings (6–8 months after surgery)
This study, to our knowledge, is the first to show that transplanted photoreceptors survive and function even with host’s dysfunctional RPE
Summary
CSC14 human embryonic stem cells were differentiated into primordial eye structures called retinal organoids. Retinal organoids were analyzed by quantitative PCR and immunofluorescence and compared with human fetal retina. A custom qPCR array (Qiagen, Germantown, MD; Cat# CAPH13339; 84 retinal genes and 5 housekeeping genes) was used to analyze the expression pattern in retinal organoids and control human fetal retinal tissues. The mean Delta Ct value per gene was determined and scatterplots of mean delta Ct values for human fetal retina vs retinal organoids was plotted. Both the “hclust” R-program algorithm to analyze qPCR data and code used to create the gene array scatterplots were downloaded from The R Foundation (https://www.r-project.org/)
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