title: From confluent human iPS cells to self-forming neural retina and retinal pigmented epithelium.
abstract: 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.
authors.affiliation: Institut de la Vision, Institut National de la Santé et de la Recherche Médicale, U968;Sorbonne Universités, Université Pierre-et-Marie-Curie Paris 6, Unité Mixte de Recherche S968;Centre National de la Recherche Scientifique, Unité Mixte de Recherche 7210, 75012 Paris, France;
authors.forename: Sacha
authors.initials: S
authors.lastname: Reichman
authors.affiliation: Institut de la Vision, Institut National de la Santé et de la Recherche Médicale, U968;Sorbonne Universités, Université Pierre-et-Marie-Curie Paris 6, Unité Mixte de Recherche S968;Centre National de la Recherche Scientifique, Unité Mixte de Recherche 7210, 75012 Paris, France;
authors.forename: Angélique
authors.initials: A
authors.lastname: Terray
authors.affiliation: Institut de la Vision, Institut National de la Santé et de la Recherche Médicale, U968;Sorbonne Universités, Université Pierre-et-Marie-Curie Paris 6, Unité Mixte de Recherche S968;Centre National de la Recherche Scientifique, Unité Mixte de Recherche 7210, 75012 Paris, France;
authors.forename: Amélie
authors.initials: A
authors.lastname: Slembrouck
authors.affiliation: Institut de la Vision, Institut National de la Santé et de la Recherche Médicale, U968;Sorbonne Universités, Université Pierre-et-Marie-Curie Paris 6, Unité Mixte de Recherche S968;Centre National de la Recherche Scientifique, Unité Mixte de Recherche 7210, 75012 Paris, France;
authors.forename: Céline
authors.initials: C
authors.lastname: Nanteau
authors.affiliation: Institut de la Vision, Institut National de la Santé et de la Recherche Médicale, U968;Sorbonne Universités, Université Pierre-et-Marie-Curie Paris 6, Unité Mixte de Recherche S968;Centre National de la Recherche Scientifique, Unité Mixte de Recherche 7210, 75012 Paris, France;
authors.forename: Gaël
authors.initials: G
authors.lastname: Orieux
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, Université d'Évry Val-d'Essonne, U861, Institute for Stem Cell Therapy, Association Française Contre les Myopathies, 91030 Evry, France; and.
authors.forename: Walter
authors.initials: W
authors.lastname: Habeler
authors.affiliation: Institut de la Vision, Institut National de la Santé et de la Recherche Médicale, U968;Sorbonne Universités, Université Pierre-et-Marie-Curie Paris 6, Unité Mixte de Recherche S968;Centre National de la Recherche Scientifique, Unité Mixte de Recherche 7210, 75012 Paris, France;
authors.forename: Emeline F
authors.initials: EF
authors.lastname: Nandrot
authors.affiliation: Institut de la Vision, Institut National de la Santé et de la Recherche Médicale, U968;Sorbonne Universités, Université Pierre-et-Marie-Curie Paris 6, Unité Mixte de Recherche S968;Centre National de la Recherche Scientifique, Unité Mixte de Recherche 7210, 75012 Paris, France;Centre Hospitalier National d'Ophtalmologie des Quinze-Vingts, Institut National de la Santé et de la Recherche Médicale-Direction de l'Hospitalisation et de l'Organisation des Soins, Centre d'Investigation Clinique 503, 75012 Paris, France.
authors.forename: José-Alain
authors.initials: JA
authors.lastname: Sahel
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, Université d'Évry Val-d'Essonne, U861, Institute for Stem Cell Therapy, Association Française Contre les Myopathies, 91030 Evry, France; and.
authors.forename: Christelle
authors.initials: C
authors.lastname: Monville
authors.affiliation: Institut de la Vision, Institut National de la Santé et de la Recherche Médicale, U968;Sorbonne Universités, Université Pierre-et-Marie-Curie Paris 6, Unité Mixte de Recherche S968;Centre National de la Recherche Scientifique, Unité Mixte de Recherche 7210, 75012 Paris, France; olivier.goureau@inserm.fr.
authors.forename: Olivier
authors.initials: O
authors.lastname: Goureau
mesh_terms: D000328:Adult; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D002478:Cells, Cultured; D020405:Dermis / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D039902:Multipotent Stem Cells / Q000166:cytology / Q000378:metabolism; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D010786:Photoreceptor Cells / Q000166:cytology / Q000378:metabolism; D055351:Retinal Neurons / Q000166:cytology* / Q000378:metabolism; D055213:Retinal Pigment Epithelium / Q000166:cytology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D018398:Homeodomain Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors
keywords: cones; retinal ganglion cells; rods