1. Isolation and Comparative Transcriptome Analysis of Human Fetal and iPSC-Derived Cone Photoreceptor Cells
- Author
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Welby, Emily, Lakowski, Jorn, Di Foggia, Valentina, Budinger, Dimitri, Gonzalez-Cordero, Anai, Lun, Aaron TL, Epstein, Michael, Patel, Aara, Cuevas, Elisa, Kruczek, Kamil, Naeem, Arifa, Minneci, Federico, Hubank, Mike, Jones, David T, Marioni, John C, Ali, Robin R, Sowden, Jane C, Lun, Aaron [0000-0002-3564-4813], Marioni, John [0000-0001-9092-0852], and Apollo - University of Cambridge Repository
- Subjects
lcsh:R5-920 ,genetic structures ,cone photoreceptor cells ,retinal organoids ,Gene Expression Profiling ,cell transplantation therapy ,Induced Pluripotent Stem Cells ,Retinal Degeneration ,Rod Opsins ,Gene Expression Regulation, Developmental ,Cell Differentiation ,eye diseases ,Article ,Retina ,retinal dystrophies ,cell surface markers ,Fetus ,lcsh:Biology (General) ,Retinal Cone Photoreceptor Cells ,Humans ,sense organs ,human pluripotent stem cells ,lcsh:Medicine (General) ,Transcriptome ,lcsh:QH301-705.5 - Abstract
Summary Loss of cone photoreceptors, crucial for daylight vision, has the greatest impact on sight in retinal degeneration. Transplantation of stem cell-derived L/M-opsin cones, which form 90% of the human cone population, could provide a feasible therapy to restore vision. However, transcriptomic similarities between fetal and stem cell-derived cones remain to be defined, in addition to development of cone cell purification strategies. Here, we report an analysis of the human L/M-opsin cone photoreceptor transcriptome using an AAV2/9.pR2.1:GFP reporter. This led to the identification of a cone-enriched gene signature, which we used to demonstrate similar gene expression between fetal and stem cell-derived cones. We then defined a cluster of differentiation marker combination that, when used for cell sorting, significantly enriches for cone photoreceptors from the fetal retina and stem cell-derived retinal organoids, respectively. These data may facilitate more efficient isolation of human stem cell-derived cones for use in clinical transplantation studies., Graphical Abstract, Highlights • Definition of an L/M-opsin cone-enriched gene signature from human fetal cones • Single-cell analysis revealed heterogeneity based on cone maturation stage • Fetal and stem cell-derived cones show similar expression of cone gene signature • FACS with SSEA1/CD133/CD26/CD147 markers enriches for human cones, Welby et al. define a cone-enriched gene signature within a human fetal L/M-opsin cone population, which is used as a baseline reference to demonstrate similar cone gene expression between bona fide and stem cell-derived L/M-opsin cone cells. Furthermore, profiling of cell surface molecules in human fetal cones led to the generation of a cluster of differentiation marker panel, which provides enrichment of fetal and stem cell-derived cones.
- Published
- 2017