Brief Report: Self-Organizing Neuroepithelium from Human Pluripotent Stem Cells Facilitates Derivation of Photoreceptors

Boucherie, Cédric; Mukherjee, Sayandip; Henckaerts, Els; Thrasher, Adrian J.; Sowden, Jane C.; Ali, Robin R.

doi:10.1002/stem.1268

Cited by 79 publications

(73 citation statements)

References 41 publications

(62 reference statements)

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“…In terms of timing, differentiation of NR-like structures is quite similar to the ones reported for the in vitro self-formed OC and OV-like structures (16,17), with the detection of RGCs between D21 and D35 and photoreceptor precursors around D30-D40 whereas more mature photoreceptors, stained for the presence of OPSIN or RHODOPSIN, were not clearly identified before D77. The use of different molecules during the culture, such as taurine, sonic hedgehog, retinoic acid, FGF1, and FGF2, could accelerate photoreceptor maturation, as recently reported for hESC aggregates in the presence of Matrigel (28). Interestingly, we report that inhibition of the Notch pathway when RPCs are committed to the photoreceptor lineage clearly enhances the proportion of photoreceptor precursors in the NR-like structures, with a twofold increase in CRX + cells, presumably achieved by forcing RPCs to exit early from the cell cycle, as previously described (16).…”

Section: Discussionsupporting

confidence: 59%

“…Early generated structures present an OV phenotype revealed by coexpression of PAX6 and RAX, and an opposite gradient of VSX2 and MITF between the neuroepithelium and the RPE. This efficiency is likely due in part to the increasing endogenous production by confluent hiPSCs of DKK1 and NOG-GIN, two inducers of neural and retinal specification, generally added for retinal differentiation of hESCS or hiPSCs (17,28).…”

Section: Discussionmentioning

confidence: 99%

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From confluent human iPS cells to self-forming neural retina and retinal pigmented epithelium

Reichman

Terray

Slembrouck

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

253

265

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Progress in retinal-cell therapy derived from human pluripotent stem cells currently faces technical challenges that require the development of easy and standardized protocols. Here, we developed a simple retinal differentiation method, based on confluent human induced pluripotent stem cells (hiPSC), bypassing embryoid body formation and the use of exogenous molecules, coating, or Matrigel. In 2 wk, we generated both retinal pigmented epithelial cells and self-forming neural retina (NR)-like structures containing retinal progenitor cells (RPCs). We report sequential differentiation from RPCs to the seven neuroretinal cell types in maturated NR-like structures as floating cultures, thereby revealing the multipotency of RPCs generated from integration-free hiPSCs. Furthermore, Notch pathway inhibition boosted the generation of photoreceptor precursor cells, crucial in establishing cell therapy strategies. This innovative process proposed here provides a readily efficient and scalable approach to produce retinal cells for regenerative medicine and for drug-screening purposes, as well as an in vitro model of human retinal development and disease.retinal ganglion cells | rods | cones I rreversible blindness caused by retinal diseases, such as inherited retinopathies, age-related macular degeneration (AMD), or glaucoma, is mainly due to the impairment or loss of function of photoreceptor cells, supporting retinal pigmented epithelium (RPE) or retinal ganglion cells (RGCs). Rescuing the degenerated retina is a major challenge for which specific cell replacement is one of the most promising approaches (1, 2). Pluripotent stem cells, like human embryonic stem cells (hESCs) or induced pluripotent stem cells (hiPSCs), have the ability to be expanded indefinitely in culture and could be used as an unlimited source of retinal cells for the treatment of retinal degenerative diseases (3, 4). Several publications have indicated that hESCs and hiPSCs can be differentiated into RPE cells spontaneously after fibroblast growth factor (FGF) 2 removal (5-7) or by different floating aggregate methods (8-11). Concerning neural retinal cells, a growing body of convergent data has demonstrated the ability of hESCs or hiPSCs to be committed into the neural retinal lineage and further differentiated into cells expressing photoreceptor markers (12-15). Recent innovative approaches using 3D -cultures from embryoid bodies (EBs) of hESCs or hiPSCs allowed the self-formation of optic cup (OC) structures (16) or the generation of optic vesicle (OV)-like structures (17), depending on the addition of exogenous molecules and different substrates used. These protocols require multiple steps and trained handling, which are not always compatible with the manufacturing process for therapeutic approach or drug screening that need a large-scale production of cells of interest. Therefore, very simple and reliable approaches minimizing the use of exogenous molecules should be developed to generate hESCs or hiPSC-derived retinal cells.In the present study, ...

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Section: Discussionsupporting

confidence: 59%

Section: Discussionmentioning

confidence: 99%

From confluent human iPS cells to self-forming neural retina and retinal pigmented epithelium

Reichman

Terray

Slembrouck

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

253

265

View full text Add to dashboard Cite

show abstract

“…23,33,36,37 Thus, one of the important tasks of transplanted hESC-RPE is to produce sufficient ECM to restore the functions of the damaged BM if no BM mimicking nonbiodegradable biomaterial is used with cells. 16,17 Previously, it has been shown that ECM affects the early stage differentiation of hPSCs into neural progenitors and neurons, 38 retinal progenitor cells, 39,40 and RPE cells. 41 In addition, different ECM proteins have been used in differentiation of hESC-RPE.…”

Section: Introductionmentioning

confidence: 99%

Structure and Barrier Properties of Human Embryonic Stem Cell–Derived Retinal Pigment Epithelial Cells Are Affected by Extracellular Matrix Protein Coating

Sorkio

Hongisto

Kaarniranta

et al. 2014

Tissue Engineering Part A

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Extracellular matrix (ECM) interactions play a vital role in cell morphology, migration, proliferation, and differentiation of cells. We investigated the role of ECM proteins on the structure and function of human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cells during their differentiation and maturation from hESCs into RPE cells in adherent differentiation cultures on several human ECM proteins found in native human Bruch's membrane, namely, collagen I, collagen IV, laminin, fibronectin, and vitronectin, as well as on commercial substrates of xeno-free CELLstartÔ and MatrigelÔ. Cell pigmentation, expression of RPE-specific proteins, fine structure, as well as the production of basal lamina by hESC-RPE on different protein coatings were evaluated after 140 days of differentiation. The integrity of hESC-RPE epithelium and barrier properties on different coatings were investigated by measuring transepithelial resistance. All coatings supported the differentiation of hESC-RPE cells as demonstrated by early onset of cell pigmentation and further maturation to RPE monolayers after enrichment. Mature RPE phenotype was verified by RPE-specific gene and protein expression, correct epithelial polarization, and phagocytic activity. Significant differences were found in the degree of RPE cell pigmentation and tightness of epithelial barrier between different coatings. Further, the thickness of self-assembled basal lamina and secretion of the key ECM proteins found in the basement membrane of the native RPE varied between hESC-RPE cultured on compared protein coatings. In conclusion, this study shows that the cell culture substrate has a major effect on the structure and basal lamina production during the differentiation and maturation of hESC-RPE potentially influencing the success of cell integrations and survival after cell transplantation.

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“…30 To avoid the presence of animal derivatives (animal-derived sera, or in most cases Matrigel), a chemically defined 3D retinal differentiation medium was recently reported, which only contains nicotinamide and 20 ng/mL of basic fibroblast growth factor. 121 To date, efforts to speed up the differentiation protocols 31,122,123 using molecules known to promote retinal differentiation 31,118,123 or by passing the EB formation 122 have generally reduced the survival of differentiated cells, 31,123 and/or the stratification obtained. 122 Whether correct lamination of pluripotent stem cell-derived donor retinal tissue is required for generation of transplantationcompetent human photoreceptor cells remains to be resolved.…”

Section: Simplicity and Speedmentioning

confidence: 99%

Induced Pluripotent Stem Cell Therapies for Degenerative Disease of the Outer Retina: Disease Modeling and Cell Replacement

Foggia

Makwana

Ali³

et al. 2016

Journal of Ocular Pharmacology and Therapeutics

Self Cite

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Stem cell therapies are being explored as potential treatments for retinal disease. How to replace neurons in a degenerated retina presents a continued challenge for the regenerative medicine field that, if achieved, could restore sight. The major issues are: (i) the source and availability of donor cells for transplantation; (ii) the differentiation of stem cells into the required retinal cells; and (iii) the delivery, integration, functionality, and survival of new cells in the host neural network. This review considers the use of induced pluripotent stem cells (iPSC), currently under intense investigation, as a platform for cell transplantation therapy. Moreover, patientspecific iPSC are being developed for autologous cell transplantation and as a tool for modeling specific retinal diseases, testing gene therapies, and drug screening.

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Brief Report: Self-Organizing Neuroepithelium from Human Pluripotent Stem Cells Facilitates Derivation of Photoreceptors

Cited by 79 publications

References 41 publications

From confluent human iPS cells to self-forming neural retina and retinal pigmented epithelium

From confluent human iPS cells to self-forming neural retina and retinal pigmented epithelium

Structure and Barrier Properties of Human Embryonic Stem Cell–Derived Retinal Pigment Epithelial Cells Are Affected by Extracellular Matrix Protein Coating

Induced Pluripotent Stem Cell Therapies for Degenerative Disease of the Outer Retina: Disease Modeling and Cell Replacement

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