Abstract
Visual impairment is a major health concern globally resulting in a severe detriment to the quality of life for patients and a global economic burden. Irreversible vision loss or blindness can result from many etiologies including optic neuropathies, developmental malformations, ocular tumors, and drug toxicities. Chief among these causes for incurable vision loss are optic neuropathies such as glaucoma, diabetic neuropathy, and traumatic optic neuropathy. These pathologies are characterized by a slow progressive loss of retinal ganglion cells (RGCs) and their axons which have negligible regenerative potential. In recent years, organoids derived from human pluripotent stem cells have emerged as a potent tool for the study and treatment of disease due to their ability to self-assemble and recapitulate embryonic tissue organization in vitro. Here, two cell lines of human induced pluripotent stem cells (hiPSC) derived from different parental cells were evaluated for their commitment to generate a retinal organoid platform. A methodology was further established to employ hiPSC-derived retinal organoids as an expandable source of electrophysiological active RGC-progenitors for regenerative ophthalmology. The use of retinal organoids derived from hiPSC not only represents a powerful tool in on-going efforts to regenerate RGCs for the treatment of vision loss but may also be used to develop more representative models of developmental ocular diseases such as retinoblastoma. Here we used CRISPR/Cas9 to establish a hiPSC cell line deficient of the retinoblastoma gene (RB1). Retinal organoids derived from this mutated cell line expressed the transcriptome profile of the different retinal cell types and the complexity that occurs during retinal development. Overall, this research established a platform of stem cell-derived retinal organoids that can serve as a source of retinal progenitor cells for regenerative ophthalmology and as an in vitro human model to study tumor formation in the developing retina.