Abstract
Human retinal organoids are three-dimensional tissue systems derived from human pluripotent stem cells (hPSCs) that recreate aspects of the cytoarchitectural organization of the retina. The potential of these retinal organoids to serve as in vitro models of retinal development and disease, as well as cell-based substrates for regenerative therapies, relies on their ability to faithfully recapitulate the cellular and functional properties of the native retina. Here, we describe a protocol allowing generation of hiPSC-derived three-dimensional retinal organoids that recapitulate spatiotemporally each of the main steps of retinal development observed in vivo and form a fully laminated retinal tissue with functional light-sensitive photoreceptors. These 3D retinal organoids contain all major retinal cell types including ganglion, amacrine, and horizontal cells; the three major bipolar cell subtypes (rod, cone ON and OFF); all photoreceptor subtypes (rods, red, green, and blue cones); and Müller cells arranged in their proper layers. Remarkably, the photoreceptors generated with this protocol are physiologically competent, form outer segments, and respond to light. Studies published in recent years demonstrate the suitability of the retinal organoids generated by this protocol as in vitro models of retinal development and disease.
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Acknowledgments
We would like to specially thank Xiufeng Zhong, Christian Gutierrez, Tian Xue, Christopher Hampton, M. Natalia Vergara, Li-Hui Cao, Ann Peters, Tea-Soon Park, Elias T. Zambidis, Jason S. Meyer, David M. Gamm, and King-Wai Yau for their contributions in establishing the protocol for generation of three-dimensional retinal tissue with functional photoreceptors from human iPSCs [9]. This work was supported by funding to M.V.C.S. from the Gates Frontiers Fund, the Solich Fund, CellSight Development Fund, and an unrestricted Research Award from Research to Prevent Blindness to the Department of Ophthalmology, University of Colorado.
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Aparicio-Domingo, S. et al. (2023). Generation of Three-Dimensional Retinal Tissue with Physiologically Competent, Light-Sensitive Photoreceptors from Human-Induced Pluripotent Stem Cells. In: Gopalakrishnan, J. (eds) Brain Organoid Research. Neuromethods, vol 189. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2720-4_6
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DOI: https://doi.org/10.1007/978-1-0716-2720-4_6
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