Robust Differentiation of mRNA-Reprogrammed Human Induced Pluripotent Stem Cells Toward a Retinal Lineage

Sridhar, Akshayalakshmi; Ohlemacher, Sarah K.; Langer, Kirstin B.; Meyer, Jason S.

doi:10.5966/sctm.2015-0093

Cited by 36 publications

(30 citation statements)

References 77 publications

(192 reference statements)

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“…Methods to generate 3D retinal cultures from hPSCs have increased in number substantially since their first descriptions (Meyer et al, 2011;Nakano et al, 2012;Phillips et al, 2012Phillips et al, , 2018bSridhar et al, 2016;Reichman et al, 2014;Zhong et al, 2014;Kuwahara et al, 2015;Mellough et al, 2015;Singh et al, 2015;Lowe et al, 2016;Wiley et al, 2016;Gonzalez-Cordero et al, 2017;Wahlin et al, 2017;Ovando-Roche et al, 2018;Hallam et al, 2018;Luo et al, 2018). Even so, most protocols follow a common developmental sequence and time frame that closely mimic normal human retinogenesis, including production of a starting population of OVs that display a phase-bright, palisading neuroepithelial rim.…”

Section: Discussionmentioning

confidence: 99%

“…The capacity to generate bona fide fetal-stage neural retinal cell types and tissues from human embryonic and induced pluripotent stem cells (hESCs and hiPSCs; collectively, hPSCs) has spurred their use in disease modeling (Tucker et al, 2011(Tucker et al, , 2013Jin et al, 2012;Phillips et al, 2014;Yoshida et al, 2015;Arno et al, 2016;Parfitt et al, 2016;Megaw et al, 2017;Schwarz et al, 2017;Sharma et al, 2017;Shimada et al, 2017;Deng et al, 2018) and photoreceptor (PR) replacement efforts (Lamba et al, 2009(Lamba et al, , 2010Hambright et al, 2012;Barnea-Cramer et al, 2016;Shirai et al, 2016;Chao et al, 2017;Gonzalez-Cordero et al, 2017;Mandai et al, 2017;Zhu et al, 2017Zhu et al, , 2018Iraha et al, 2018;Lakowski et al, 2018;McLelland et al, 2018;Gagliardi et al, 2018). Most of these studies employ hPSC differentiation methods that propagate retinal progeny as isolated 3D optic vesicle-like structures (OVs), also known as retinal organoids, in suspension culture (Meyer et al, 2009(Meyer et al, , 2011Nakano et al, 2012;Phillips et al, 2012Phillips et al, , 2018bSridhar et al, 2016;Reichman et al, 2014;Zhong et al, 2014;Kuwahara et al, 2015;Mellough et al, 2015;Singh et al, 2015;Lowe et al, 2016;…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2018

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

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

et al. 2018

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“…hPSCs were maintained as previously described . Briefly, three lines of control human pluripotent stem cells (H9, H7 and miPS2 ) were used, and three lines of patient‐derived induced pluripotent stem cells from an OPTN E50K patient were derived. All cell lines were maintained in the pluripotent state with mTeSR1 medium (Stemcell Technologies, Vancouver Canada, http://www.stemcell.com) on matrigel‐coated six‐well plates.…”

Section: Methodsmentioning

confidence: 99%

Stepwise Differentiation of Retinal Ganglion Cells from Human Pluripotent Stem Cells Enables Analysis of Glaucomatous Neurodegeneration

et al. 2016

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Human pluripotent stem cells (hPSCs), including both embryonic and induced pluripotent stem cells, possess the unique ability to readily differentiate into any cell type of the body, including cells of the retina. Although previous studies have demonstrated the ability to differentiate hPSCs to a retinal lineage, the ability to derive retinal ganglion cells (RGCs) from hPSCs has been complicated by the lack of specific markers with which to identify these cells from a pluripotent source. In the current study, the definitive identification of hPSC-derived RGCs was accomplished by their directed, stepwise differentiation through an enriched retinal progenitor intermediary, with resultant RGCs expressing a full complement of associated features and proper functional characteristics. These results served as the basis for the establishment of induced pluripotent stem cells (iPSCs) from a patient with a genetically inherited form of glaucoma, which results in damage and loss of RGCs. Patient-derived RGCs specifically exhibited a dramatic increase in apoptosis, similar to the targeted loss of RGCs in glaucoma, which was significantly rescued by the addition of candidate neuroprotective factors. Thus, the current study serves to establish a method by which to definitively acquire and identify RGCs from hPSCs and demonstrates the ability of hPSCs to serve as an effective in vitro model of disease progression. Moreover, iPSC-derived RGCs can be utilized for future drug screening approaches to identify targets for the treatment of glaucoma and other optic neuropathies.

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“…For unknown reasons, retina organoids cultured by different groups gain light sensitivity only *15% of the time. 6,8,9 A recent study optimized culture conditions to produce more mature light-sensitive tissue in a scalable manner by optimizing both cell seeding density and culture nutrient condition. 6 When organoids were cultured individually in 96-well plates, these showed a marked increase in the percentage of organoids that exhibited light sensitivity by electrophysiological testing; retina organoids cultured in large groups in 6-well plates were sensitive to light *25% of the time, but *37% of organoids cultured individually were light sensitive.…”

Section: Recent Advancesmentioning

confidence: 99%

Improved Ocular Tissue Models and Eye-On-A-Chip Technologies Will Facilitate Ophthalmic Drug Development

Wright

Becker

Low

et al. 2020

Journal of Ocular Pharmacology and Therapeutics

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In this study, we describe efforts by the National Eye Institute (NEI) and National Center for Advancing Translational Science (NCATS) to catalyze advances in 3-dimensional (3-D) ocular organoid and microphysiological systems ( MPS). We reviewed the recent literature regarding ocular organoids and tissue chips. Animal models, 2-dimensional cell culture models, and postmortem human tissue samples provide the vision research community with insights critical to understanding pathophysiology and therapeutic development. The advent of induced pluripotent stem cell technologies provide researchers with enticing new approaches and tools that augment study in more traditional models to provide the scientific community with insights that have previously been impossible to obtain. Efforts by the National Institutes of Health (NIH) have already accelerated the pace of scientific discovery, and recent advances in ocular organoid and MPS modeling approaches have opened new avenues of investigation. In addition to more closely recapitulating the morphologies and physiological responses of in vivo human tissue, key breakthroughs have been made in the past year to resolve long-standing scientific questions regarding tissue development, molecular signaling, and pathophysiological mechanisms that promise to provide advances critical to therapeutic development and patient care. 3-D tissue culture modeling and MPS offer platforms for future high-throughput testing of therapeutic candidates and studies of gene interactions to improve models of complex genetic diseases with no well-defined etiology, such as age-related macular degeneration and Fuchs' dystrophy.

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Robust Differentiation of mRNA-Reprogrammed Human Induced Pluripotent Stem Cells Toward a Retinal Lineage

Cited by 36 publications

References 77 publications

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

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

Stepwise Differentiation of Retinal Ganglion Cells from Human Pluripotent Stem Cells Enables Analysis of Glaucomatous Neurodegeneration

Improved Ocular Tissue Models and Eye-On-A-Chip Technologies Will Facilitate Ophthalmic Drug Development

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