Abstract
BackgroundDysfunction of the retinal pigment epithelium (RPE) is implicated in numerous forms of retinal degeneration. The readily accessible environment of the eye makes it particularly suitable for the transplantation of RPE cells, which can now be derived from autologous induced pluripotent stem cells (iPSCs), to treat retinal degeneration. For RPE transplantation to become feasible in the clinic, patient-specific somatic cells should be reprogrammed to iPSCs without the introduction of reprogramming genes into the genome of the host cell, and then subsequently differentiated into RPE cells that are well characterized for safety and functionality prior to transplantation.MethodsWe have reprogrammed human dermal fibroblasts to iPSCs using nonintegrating RNA, and differentiated the iPSCs toward an RPE fate (iPSC-RPE), under Good Manufacturing Practice (GMP)-compatible conditions.ResultsUsing highly sensitive assays for cell polarity, structure, organelle trafficking, and function, we found that iPSC-RPE cells in culture exhibited key characteristics of native RPE. Importantly, we demonstrate for the first time with any stem cell-derived RPE cell that live cells are able to support dynamic organelle transport. This highly sensitive test is critical for RPE cells intended for transplantation, since defects in intracellular motility have been shown to promote RPE pathogenesis akin to that found in macular degeneration. To test their capabilities for in-vivo transplantation, we injected the iPSC-RPE cells into the subretinal space of a mouse model of retinal degeneration, and demonstrated that the transplanted cells are capable of rescuing lost RPE function.ConclusionsThis report documents the successful generation, under GMP-compatible conditions, of human iPSC-RPE cells that possess specific characteristics of healthy RPE. The report adds to a growing literature on the utility of human iPSC-RPE cells for cell culture investigations on pathogenicity and for therapeutic transplantation, by corroborating findings of others, and providing important new information on essential RPE cell biological properties.
Highlights
Dysfunction of the retinal pigment epithelium (RPE) is implicated in numerous forms of retinal degeneration
This paper reports the use of a nonintegrating approach to generate Induced pluripotent stem cell (iPSC) for the generation of RPE cells under Good Manufacturing Practice (GMP)-compatible practices
Our results show that iPSCs, generated with a nonintegrating method, can serve as a renewable source of functional RPE cells, which can be used for detailed cell biological analyses of pathogenicity in vitro, as well as for transplantation in treatment of retinal diseases
Summary
Dysfunction of the retinal pigment epithelium (RPE) is implicated in numerous forms of retinal degeneration. The readily accessible environment of the eye makes it suitable for the transplantation of RPE cells, which can be derived from autologous induced pluripotent stem cells (iPSCs), to treat retinal degeneration. One promising form of treatment for AMD is the transplantation of healthy RPE cells into the retinas of human patients to restore lost functions, and potentially halt or reverse the progression of the disease. Patient-derived iPSC-RPE cells offer disease modeling and testing of pharmacologically active compounds, in addition to autologous transplantation, without the need for immunosuppression [6]. The current technology allows for the generation of patient-specific iPSCs that are free of integrated reprogramming genes, and can subsequently be used to generate the quantities of functional RPE cells necessary for transplantation purposes
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