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Single-cell RNA-seq reveals the molecular progression of retinal progenitors in human embryonic stem cell models of early retinogenesis

ABSTRACT: We used single-cell RNA-sequencing (scRNA-seq) on human embryonic stem cell (hESC)-derived retinal organoid to deconstruct RPC developmental trajectory in the early retinogenesis, which revealed the fate branchpoint in RPC progression.

ORGANISM(S): Homo sapiens

PROVIDER: GSE122783 | GEO | 2019/08/22

REPOSITORIES: GEO

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Bankable human iPSC-derived Retinal Progenitors as a New Source of Multipotent Cells

Project description:Retinal progenitor cells (RPCs) are the source of all retinal cell types during retinogenesis. Until now, the isolation and expansion of RPCs has been at the expense of their multipotency. Here, we report simple methods and media for the generation, expansion, and cryopreservation of human induced pluripotent stem-cell derived-RPCs (hiRPCs). Thawed and passed hiRPCs maintained biochemical and transcriptional RPC phenotypes and their ability to differentiate into all retinal cell types. Specific conditions allowed the generation of large cultures of photoreceptor precursors enriched up to 90% within a few weeks and without a purification step. Combined RNA-seq analysis between hiRPCs and retinal organoids identified genes involved in developmental or degenerative retinal diseases. Thus, hiRPC lines could provide a new source of retinal cells for cell-based therapies or drug discovery and could be an advanced cellular tool to better understand retinal dystrophies.

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Retinal Cell Type Epigenetic Memory Predicts Reprogramming Efficiency and Retinogenesis in 3D Organoid Cultures [TEBS_Mm]

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Retinal Cell Type Epigenetic Memory Predicts Reprogramming Efficiency and Retinogenesis in 3D Organoid Cultures [RNA-Seq_Mm]

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Project description:To study the function of Mettl3 during retinal development, we used Six3-Cre and Mettl3floxed mice to conditionally knock out Mettl3 from retinal progenitor cells. The mutant mice show defects in late-stage retinogenesis and structural and physiological homeostasis of the retina.

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Transcriptomic analysis of progenitor marker expression in retinal cells

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DNA methylation in hESC derived brain organoids and iPSC derived neurons, astrocytes and neural progenitors

Project description:We studied the impact of ZIKA virus infection on DNA methylation in whole organoids, organoid derived astrocytes, neurons neural progeniotors and hESC or iPSC derived astrocytes, neurons, neural progenotor cells

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Project description:The plasticity of pluripotent stem cells provides new possibilities for studying development, degeneration, and regeneration. Protocols for the differentiation of retinal organoids from embryonic stem cells have been developed, which either recapitulate complete eyecup morphogenesis or maximize photoreceptor genesis. Here, we have developed a protocol for the efficient generation of large, 3D-stratified retinal organoids that does not require evagination of optic-vesicle-like structures, which so far limited the organoid yield. Analysis of gene expression in individual organoids, cell birthdating, and interorganoid variation indicate efficient, reproducible, and temporally regulated retinogenesis. Comparative analysis of a transgenic reporter for PAX6, a master regulator of retinogenesis, shows expression in similar cell types in mouse in vivo, and in mouse and human retinal organoids. Early or late Notch signaling inhibition forces cell differentiation, generating organoids enriched with cone or rod photoreceptors, respectively, demonstrating the power of our improved organoid system for future research in stem cell biology and regenerative medicine.

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