Accelerated photoreceptor differentiation of hiPSC-derived retinal organoids by contact co-culture with retinal pigment epithelium

Akhtar, Tashfeen; Xie, Haohuan; Khan, Muhammad Imran; Zhao, Huan; Bao, Jin; Zhang, Mei; Xue, Tian

doi:10.1016/j.scr.2019.101491

Cited by 52 publications

(41 citation statements)

References 37 publications

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“…It quickly became evident that once the self-formation of retinal organoids is done (which can be achieved with a number of protocols maintaining and promoting the propensity of retina (the outpocketing of anterior neuroectoderm) to form, further maturation and long-term maintenance of retina-in-a-dish requires more sophisticated media providing more salts, anti-oxidants, a milieu of supporting paracrine factors (provided by serum or serum plus RPE conditioned medium), etc. ( Zhong et al, 2014 ; Bardy et al, 2015 ; Singh et al, 2015 ; Di Lauro et al, 2016 ; Achberger et al, 2019 ; Akhtar et al, 2019 ; Capowski et al, 2019 ). These issues are mostly technical, and current biomanufacturing technologies allow generating large number of aggregates of a defined size (e.g., for large-scale drug discovery) if needed.…”

Section: Lack Of Rpe Photoreceptor Interaction In Retinal Organoidsmentioning

confidence: 99%

“…Pigment epithelium derived factor (PEDF, or SERPINF1) signaling is an important paracrine and autocrine pathway in retina ( Tombran-Tink et al, 1991 ; Malchiodi-Albedi et al, 1998 ; Tombran-Tink and Barnstable, 2003 ) for maintaining PR-RPE niche ( Volpert et al, 2009 ; Akiyama et al, 2012 ), photoreceptor maturation ( Jablonski et al, 2000 ; Akhtar et al, 2019 ) and survival ( Comitato et al, 2018 ; Chen Y. et al, 2019 ). PEDF expression is a hallmark of RPE maturation and polarization ( Strunnikova et al, 2010 ; Maruotti et al, 2015 ; McGill et al, 2017 ).…”

Section: Signaling Pathways Involved In Human Retinal Development Dementioning

confidence: 99%

“…However, some in vivo results reveal better organization of OSs and longer OSs in the long-term subretinal grafts ( Shirai et al, 2016 ), all pointing toward the lumen of rosette-like photoreceptor aggregates in subretinal space ( Shirai et al, 2016 ; Nasonkin et al, 2019 ). Naturally, there is no ongoing renewal and phagocytosis processes in long term retinal cell and retinal organoid cultures, but it has been expected for a long while that with developing photoreceptor-RPE co-culture systems faithfully recapitulating structure and function of the subretinal niche, OS elongation and photoreceptor-RPE biology can be reestablished ( German et al, 2008 ; Di Lauro et al, 2016 ; DiStefano et al, 2018 ; Akhtar et al, 2019 ; Brooks et al, 2019 ; Capowski et al, 2019 ). As work on retinal repair rapidly progresses ( Holmes, 2018a , b ; Makin, 2019 ), these technologies will likely be developed in the next 3–5 years as a cheaper and robust model for drug screening and discovery, as well as platform for biomanufacturing 3D retinal tissue transplants to repair vision in advanced RD patients.…”

Section: Lack Of Rpe Photoreceptor Interaction In Retinal Organoidsmentioning

confidence: 99%

“…This could be one of many factors causing gradual degeneration of photoreceptors in long-term organoid cultures. It is expected that recreation of photoreceptor-RPE niche in a dish would make it possible to substantially increase the viability of photoreceptors in long-term in vitro cultures ( Di Lauro et al, 2016 ; Achberger et al, 2019 ; Akhtar et al, 2019 ; Chen Y. et al, 2019 ), thus enabling disease modeling and drug screening of diseases, where the integrity of subretinal niche and photoreceptor-RPE structural and functional connectivity is of paramount importance for maintaining visual function.…”

Section: Lack Of Rpe Photoreceptor Interaction In Retinal Organoidsmentioning

confidence: 99%

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Limitations and Promise of Retinal Tissue From Human Pluripotent Stem Cells for Developing Therapies of Blindness

Singh

Nasonkin

2020

Front. Cell. Neurosci.

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The self-formation of retinal tissue from pluripotent stem cells generated a tremendous promise for developing new therapies of retinal degenerative diseases, which previously seemed unattainable. Together with use of induced pluripotent stem cells or/and CRISPR-based recombineering the retinal organoid technology provided an avenue for developing models of human retinal degenerative diseases “in a dish” for studying the pathology, delineating the mechanisms and also establishing a platform for large-scale drug screening. At the same time, retinal organoids, highly resembling developing human fetal retinal tissue, are viewed as source of multipotential retinal progenitors, young photoreceptors and just the whole retinal tissue, which may be transplanted into the subretinal space with a goal of replacing patient’s degenerated retina with a new retinal “patch.” Both approaches (transplantation and modeling/drug screening) were projected when Yoshiki Sasai demonstrated the feasibility of deriving mammalian retinal tissue from pluripotent stem cells, and generated a lot of excitement. With further work and testing of both approaches in vitro and in vivo , a major implicit limitation has become apparent pretty quickly: the absence of the uniform layer of Retinal Pigment Epithelium (RPE) cells, which is normally present in mammalian retina, surrounds photoreceptor layer and develops and matures first. The RPE layer polarize into apical and basal sides during development and establish microvilli on the apical side, interacting with photoreceptors, nurturing photoreceptor outer segments and participating in the visual cycle by recycling 11-trans retinal (bleached pigment) back to 11-cis retinal. Retinal organoids, however, either do not have RPE layer or carry patches of RPE mostly on one side, thus directly exposing most photoreceptors in the developing organoids to neural medium. Recreation of the critical retinal niche between the apical RPE and photoreceptors, where many retinal disease mechanisms originate, is so far unattainable, imposes clear limitations on both modeling/drug screening and transplantation approaches and is a focus of investigation in many labs. Here we dissect different retinal degenerative diseases and analyze how and where retinal organoid technology can contribute the most to developing therapies even with a current limitation and absence of long and functional outer segments, supported by RPE.

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Section: Lack Of Rpe Photoreceptor Interaction In Retinal Organoidsmentioning

confidence: 99%

Section: Signaling Pathways Involved In Human Retinal Development Dementioning

confidence: 99%

Section: Lack Of Rpe Photoreceptor Interaction In Retinal Organoidsmentioning

confidence: 99%

Section: Lack Of Rpe Photoreceptor Interaction In Retinal Organoidsmentioning

confidence: 99%

See 2 more Smart Citations

Limitations and Promise of Retinal Tissue From Human Pluripotent Stem Cells for Developing Therapies of Blindness

Singh

Nasonkin

2020

Front. Cell. Neurosci.

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“…It is well known that RPE cell contact or secretions, such as pigment epithelium-derived factor, affect the neurosensory retina. [16][17][18][19] Recent studies have demonstrated the importance of cell-cell contact in fostering the differentiation of human retinal organoids. 18 The retinal organoids were seeded on top of a monolayer of primary cultures of RPE that were isolated from mice.…”

mentioning

confidence: 99%

Partially Differentiated Neuroretinal Cells Promote Maturation of the Retinal Pigment Epithelium

Singh

Chen

Xia

et al. 2020

Invest. Ophthalmol. Vis. Sci.

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PURPOSE. Many studies have demonstrated the ability of the retinal pigment epithelium (RPE) to foster the maturation of the developing retina. Few studies have examined the reciprocal effects of developing retina on the RPE. METHODS. RPE isolated from human fetal RPE or differentiated from human stem cells was cultured on Transwell filter inserts. Retinal progenitor cells (RPCs) were differentiated from human stem cells and cultured on a planar scaffold composed of gelatin, chondroitin sulfate, hyaluronic acid, and laminin-521. Cultures were analyzed by quantitative RT-PCR, immunofluorescence, immunoblotting, and transepithelial electrical resistance (TER). RESULTS. RPCs initially differentiated into several retina-like cell types that segregated from one another and formed loosely organized layers or zones. With time, the presumptive photoreceptor and ganglion cell layers persisted, but the intervening zone became dominated by cells that expressed glial markers with no evidence of bipolar cells or interneurons. Co-culture of this underdeveloped retinoid with the RPE resulted in a thickened layer of recoverin-positive cells but did not prevent the loss of interneuron markers in the intervening zone. Although photoreceptor inner and outer segments were not observed, immunoblots revealed that co-culture increased expression of rhodopsin and red/green opsin. Co-culture of the RPE with this underdeveloped retinal culture increased the TER of the RPE and the expression of RPE signature genes. CONCLUSIONS. These studies indicated that an immature neurosensory retina can foster maturation of the RPE; however, the ability of RPE alone to foster maturation of the neurosensory retina is limited.

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Microfluidic Techniques for Next‐Generation Organoid Systems

Gong

Kang

et al. 2022

Adv Materials Inter

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PSCs (iPSCs) and embryonic stem cells (ESCs), or adult stem cells (ASCs). Pioneered in the Yoshiki Sasai lab, where cortical tissues and optic-cuplike structures were firstly induced from ESCs in vitro, [1,2] organoid technology is regarded as a milestone in the stem cell field because it cast light upon the path to stably growing self-aggregating and selfrenewing stem cells into native tissues by mimicking developmental processes. [3] Subsequently, the rapidly developing field of organoid technology is advancing the maturity of technology for developing 3D macrostructures of organs such as the retina, [4] heart, [5] liver, [6] kidney, [7] and intestines. [8] In marked contrast to traditional 2D monocultures of cell lines losing cell-cell and cell-matrix interactions, organoids maintain and define in situ phenotypes fairly well. Despite organoids having shown great progress in tissue morphogenesis and organogenesis, the lack of organ-specific biochemical and physical microenvironments and nonphysiological shape and size have limited the function and real-life applications of organoids. Therefore, innovative strategies are urgently needed to precisely control perfusion conditions, nutrient supply, and mechanical cues in order to establish an optimal microenvironment for organoid cultures.The recently developed organs-on-a-chip (OOC) technology provides an in vivo-like microenvironment, showing great potential to enhance our understanding of organ development, physiology, and pathology. [9] Besides, the application of dynamic flow conditions or mechanical stimuli provided by OOCs can improve the terminal maturation of cells. [10,11] OOCs usually are comprised of specialized cell types and tissue elements (biochemical and biophysical cues of in vivo microenvironment), aiming to capture key functions of a specific organ. [12] Organoids rely on the self-organization of growing stem cell aggregates, whereas OOC is based on a reductionist engineering approach. In addition, organoids are multicellular 3D tissue with an architecture resembling some aspects of original tissues, which are not classified as OOC because of their production through stochastic self-organization from PSCs or ASCs and lack of microenvironment elements. [13] However, organoid and OOC techniques each have their own advantages, and they can be appropriately combined to enable highfidelity human modeling from stem cells. [14] Such synergistic Organoids are 3D multicellular structures derived from pluripotent stem cells (PSCs) or adult stem cells (ASCs), which have attracted increasing interest in the fields of drug screening, cell therapy, and regenerative medicine. Despite considerable success in culturing organoids with native microanatomy, challenges to achieving a physiologically relevant microenvironment remain. Complex dynamic feedback between cells and the extracellular matrix and uncontrollable mechano-physiological cues hamper the further study of organoid systems. Innovative engineering approaches are needed to produce, control, and analy...

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Accelerated photoreceptor differentiation of hiPSC-derived retinal organoids by contact co-culture with retinal pigment epithelium

Cited by 52 publications

References 37 publications

Limitations and Promise of Retinal Tissue From Human Pluripotent Stem Cells for Developing Therapies of Blindness

Limitations and Promise of Retinal Tissue From Human Pluripotent Stem Cells for Developing Therapies of Blindness

Partially Differentiated Neuroretinal Cells Promote Maturation of the Retinal Pigment Epithelium

Microfluidic Techniques for Next‐Generation Organoid Systems

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