Abstract
Embryonic and induced pluripotent stem cells (IPSCs) derived from mammalian species are valuable tools for modeling human disease, including retinal degenerative eye diseases that result in visual loss. Restoration of vision has focused on transplantation of neural progenitor cells (NPCs) and retinal pigmented epithelium (RPE) to the retina. Here we used transgenic common marmoset (Callithrix jacchus) and human pluripotent stem cells carrying the enhanced green fluorescent protein (eGFP) reporter as a model system for retinal differentiation. Using suspension and subsequent adherent differentiation cultures, we observed spontaneous in vitro differentiation that included NPCs and cells with pigment granules characteristic of differentiated RPE. Retinal cells derived from human and common marmoset pluripotent stem cells provide potentially unlimited cell sources for testing safety and immune compatibility following autologous or allogeneic transplantation using nonhuman primates in early translational applications.
Highlights
Novel applications of stem-cell-based therapies have revolutionized how degenerative diseases are approached
Previous studies have demonstrated the capacity of human embryonic stem cells (HESCs) and human-induced pluripotent stem cells (HIPSCs) to differentiate into cells with retinal pigmented epithelium (RPE) morphology, function, and molecular phenotypes [4, 5]
Foreskin fibroblast cells (ATCC) were propagated in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 1 mM Glutamax-I, and 1 mM nonessential amino acid (NEAA). 293FT cells were used as a packaging, cell line for generating retroviruses. 293FT were transfected with FuGENE HD with pMXS-OCT4, -SOX2 or -KLF4 plasmid, pHIT60 packaging and pVSV-G envelope construct
Summary
Novel applications of stem-cell-based therapies have revolutionized how degenerative diseases are approached. Given the propensity of stem cells to differentiate to neuronal pathways, diseases affecting the nervous system and associated tissues, such as the retina, are of great value. Retinal diseases, such as age-related macular degeneration (AMD), retinitis pigmentosa, and Stargardt disease, that render individuals functionally blind are commonly the result of impaired or complete loss of function of the photoreceptor cells or supporting retinal pigmented epithelium (RPE) [1,2,3]. HESC-, HIPSC- and fetal-derived RPE have been used to study the extent to which transplantation can correct retinal degenerative diseases [2, 5]. Preclinical studies in dystrophic rats have reported the ability of HESC-derived RPE cells to rescue visual function [1]
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