Reproducibility and staging of 3D human retinal organoids across multiple pluripotent stem cell lines

Development, Год журнала: 2018, Номер unknown

Опубликована: Янв. 1, 2018

Numerous protocols have been described that produce neural retina from human pluripotent stem cells (hPSCs), many of which are based on the culture 3D organoids. While nearly all such methods yield at least partial segments highly mature-appearing retinal structure, variabilities exist within and between organoids can change over a protracted time course differentiation. Adding to this complexity potential differences in composition configuration when viewed across multiple differentiations hPSC lines. In an effort better understand current capabilities limitations these cultures, we generated 16 lines monitored their appearance structural organization by light microscopy, immunocytochemistry, metabolic imaging, electron microscopy. We also employed optical coherence tomography imaging techniques assess compare whole or broad regions avoid selection bias. Results study led development practical staging system reduce inconsistencies organoid cultures increase rigor utilizing them developmental studies, disease modeling transplantation.

Язык: Английский

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Single-Cell Transcriptomic Comparison of Human Fetal Retina, hPSC-Derived Retinal Organoids, and Long-Term Retinal Cultures

Cell Reports, Год журнала: 2020, Номер 30(5), С. 1644 - 1659.e4

Опубликована: Фев. 1, 2020

Highlights•scRNA-seq of human retina highlights key developmental transition states•hPSC-derived retinal organoids mimic fetal cellular composition•Inner layers in exhibit differences gene expression and organization•Fetal cultures resemble but maintain better inner laminationSummaryTo study the development retina, we use single-cell RNA sequencing (RNA-seq) at stages follow major cell types as well populations transitional cells. We also analyze stem (hPSC)-derived organoids; although have a very similar composition equivalent ages there are some particular types. Moreover, lamination is disrupted more advanced compared with retina. To determine whether disorganization due to culture conditions, maintained under conditions. These retinospheres develop for least 6 months, displaying than organoids. Our (scRNA-seq) comparisons organoids, provide resource developing vitro models disease.Graphical abstract

Язык: Английский

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Stemming retinal regeneration with pluripotent stem cells

Progress in Retinal and Eye Research, Год журнала: 2018, Номер 69, С. 38 - 56

Опубликована: Ноя. 9, 2018

Cell replacement therapy is a promising treatment for irreversible retinal cell death in diverse diseases, such as age-related macular degeneration (AMD), Stargardt's disease, retinitis pigmentosa (RP) and glaucoma. These diseases are all characterized by the of one or two types that cannot regenerate spontaneously humans. Aberrant pigment epithelial (RPE) cells can be observed through optical coherence tomography (OCT) AMD patients. In RP patients, morphological functional abnormalities RPE photoreceptor layers caused genetic abnormality. disease juvenile degeneration, which loss photoreceptors area, causes central vision at an early age. Loss ganglion (RGCs) patients with Once triggered, no treatments reverse it. Transplantation-based approaches have been proposed universal to target various concomitant diseases. Both dead neuroprotection strategies used rescue visual function animal models degeneration. Diverse derived from pluripotent stem cells, including photoreceptors, RGCs even organoids layered structure, provide unlimited sources transplantation. addition, mesenchymal (MSCs) multifunctional protect degenerating cells. The aim this review summarize current findings preclinical clinical studies. We begin brief introduction degenerative followed methods generation. Preclinical studies discussed, future concerns about efficacy, safety immunorejection also addressed.

Язык: Английский

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Advancing cell therapy for neurodegenerative diseases

Cell stem cell, Год журнала: 2023, Номер 30(5), С. 512 - 529

Опубликована: Апрель 20, 2023

Язык: Английский

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Three-Dimensional Retinal Organoids Facilitate the Investigation of Retinal Ganglion Cell Development, Organization and Neurite Outgrowth from Human Pluripotent Stem Cells

Scientific Reports, Год журнала: 2018, Номер 8(1)

Опубликована: Сен. 24, 2018

Retinal organoids are three-dimensional structures derived from human pluripotent stem cells (hPSCs) which recapitulate the spatial and temporal differentiation of retina, serving as effective in vitro models retinal development. However, a lack emphasis has been placed upon development organization ganglion (RGCs) within organoids. Thus, initial efforts were made to characterize RGC throughout early stages organoid development, with clearly defined layer developing temporally-appropriate manner expressing complement RGC-associated markers. Beyond studies may also prove useful for cellular replacement extensive axonal outgrowth is necessary reach post-synaptic targets. Organoid-derived RGCs could help elucidate factors promoting outgrowth, thereby identifying approaches circumvent formidable obstacle replacement. As such, additional demonstrated significant enhancement neurite through modulation both substrate composition growth factor signaling. Additionally, organoid-derived exhibited diverse phenotypes, extending elaborate cones numerous guidance receptors. Collectively, these results establish valuable tool demonstrate utility an platform study influencing RGCs.

Язык: Английский

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Pluripotent stem cell-derived retinal organoids for disease modeling and development of therapies

Stem Cells, Год журнала: 2020, Номер 38(10), С. 1206 - 1215

Опубликована: Июнь 7, 2020

Abstract Retinal diseases constitute a genetically and phenotypically diverse group of clinical conditions leading to vision impairment or blindness with limited treatment options. Advances in reprogramming somatic cells induced pluripotent stem generation three-dimensional organoids resembling the native retina offer promising tools interrogate disease mechanisms evaluate potential therapies for currently incurable retinal neurodegeneration. Next-generation sequencing, single-cell analysis, advanced electrophysiology, high-throughput screening approaches are expected greatly expand utility cell-derived developing personalized treatments. In this review, we discuss current status future combining as human models recent technologies advance development gene, cell, drug retinopathies.

Язык: Английский

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Deconstructing Retinal Organoids: Single Cell RNA-Seq Reveals the Cellular Components of Human Pluripotent Stem Cell-Derived Retina

Stem Cells, Год журнала: 2018, Номер 37(5), С. 593 - 598

Опубликована: Дек. 12, 2018

Abstract The rapid improvements in single cell sequencing technologies and analyses afford greater scope for dissecting organoid cultures composed of multiple types create an opportunity to interrogate these models understand tissue biology, cellular behavior interactions. To this end, retinal organoids generated from human embryonic stem cells (hESCs) were analyzed by RNA-sequencing (scRNA-Seq) at three time points differentiation. Combinatorial data all revealed the presence nine clusters, five which corresponded key types: pigment epithelium (RPE), ganglion (RGCs), cone rod photoreceptors, Müller glia. remaining four clusters expressed genes typical mitotic cells, extracellular matrix components those involved homeostasis. clustering analysis decreasing RGCs, formation a distinct RPE cluster, emergence photoreceptors photoreceptor precursors, increasing number glia over time. Pseudo-time resembled order birth during development, with cluster commencing trajectory large majority completing line. Together, demonstrate feasibility potential scRNA-Seq dissect inherent complexity orderly types. Stem Cells 2019;37:593–598

Язык: Английский

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Neuroprotective Strategies for Retinal Ganglion Cell Degeneration: Current Status and Challenges Ahead

International Journal of Molecular Sciences, Год журнала: 2020, Номер 21(7), С. 2262 - 2262

Опубликована: Март 25, 2020

The retinal ganglion cells (RGCs) are the output of retina into brain. In mammals, these not able to regenerate their axons after optic nerve injury, leaving patients with neuropathies permanent visual loss. An effective RGCs-directed therapy could provide a beneficial effect prevent progression disease. Axonal injury leads functional loss RGCs and subsequently induces neuronal death, axonal regeneration would be essential restore connectivity, reestablish function system. manipulation several intrinsic extrinsic factors has been proposed in order stimulate repairing connections pathway. However, there is missing point process since, until now, no therapeutic strategy directed promote as approach for neuropathies.

Язык: Английский

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Retinal Ganglion Cells—Diversity of Cell Types and Clinical Relevance

Frontiers in Neurology, Год журнала: 2021, Номер 12

Опубликована: Май 21, 2021

Retinal ganglion cells (RGCs) are the bridging neurons that connect retinal input to visual processing centres within central nervous system. There is a remarkable diversity of RGCs and various subtypes have unique morphological features, distinct functions, characteristic pathways linking inner retina relevant brain areas. A number psychophysical electrophysiological tests been refined investigate this large varied population RGCs. Technological advances, such as high-resolution optical coherence tomography imaging, provided additional tools define pattern RGC involvement chronological sequence events in both inherited acquired optic neuropathies. The mechanistic insights gained from these studies, particular selective vulnerability relative resilience subtypes, fundamental importance they directly development targeted therapies for invariably progressive blinding diseases. This review provides comprehensive description types RGCs, developments proposed methods classification, current gaps our knowledge how differentially affected depending on underlying aetiology. synthesis body potentially amenable therapeutic modulation will hopefully lead much needed effective treatments patients with

Язык: Английский

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Role of Oxidative Stress in Ocular Diseases Associated with Retinal Ganglion Cells Degeneration

Antioxidants, Год журнала: 2021, Номер 10(12), С. 1948 - 1948

Опубликована: Дек. 5, 2021

Ocular diseases associated with retinal ganglion cell (RGC) degeneration is the most common neurodegenerative disorder that causes irreversible blindness worldwide. It characterized by visual field defects and progressive optic nerve atrophy. The underlying pathophysiology mechanisms of RGC in several ocular remain largely unknown. RGCs are a population central nervous system neurons, their soma located retina long axons extend through to form distal terminals connections brain. Because this unique cytoarchitecture highly compartmentalized energy demand, mitochondrial-dependent for adenosine triphosphate (ATP) production. Recently, oxidative stress mitochondrial dysfunction have been found be principal as well other disorders. Here, we review role degenerations, including glaucoma, hereditary atrophy, inflammatory neuritis, ischemic neuropathy, traumatic drug toxicity. We also experimental approaches using animal models research on degeneration. Lastly, discuss application antioxidants potential future therapy degenerations.

Язык: Английский

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