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CNRS, UMR_7210

6 ARTICLES PUBLISHED IN JoVE

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Medicine

Transcriptomic Analysis of Human Retinal Surgical Specimens Using jouRNAl
Marie-Noëlle Delyfer 1,2,3,4, Najate Aït-Ali 1,2,3, Hawa Camara 1,2,3, Emmanuelle Clérin 1,2,3, Jean-François Korobelnik 4, José-Alain Sahel 1,2,3, Thierry Léveillard 1,2,3
1U968, Institut National de la Santé et de la Recherche Médicale, 2UMR S 968, Université Pierre et Marie Curie, 3UMR 7210, Centre National de la Recherche Scientifique, 4Départment d'Ophtalmologie, Centre Hospitalier Universitaire de Bordeaux

We used retinal samples from retinectomy for a transcriptomic analysis of retinal detachment. We developed a procedure that allows RNA conservation between the surgical blocks and the laboratory. We standardized a protocol to purify RNA by cesium chloride ultracentrifugation to assure that the purified RNAs are suitable for microarray analysis.

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Neuroscience

Vibratome Sectioning Mouse Retina to Prepare Photoreceptor Cultures
Emmanuelle Clérin 1,4,5,6, Ying Yang 1,4,5,6, Valérie Forster 2,4,5,6, Valérie Fontaine 3,4,5,6, José-Alain Sahel 4,5,6, Thierry Léveillard 1,4,5,6
1Department of Genetics, UMR_S 968, Institut de la Vision, 2Department of Visual Information, UMR_S 968, Institut de la Vision, 3Exploratory Team, UMR_S 968, Institut de la Vision, 4Sorbonne Universités, Paris 06, UMR_S 968, Institut de la Vision, 5INSERM, U968, Institut de la Vision, 6CNRS, UMR_7210, Institut de la Vision

Neural retina of a mouse aged 8 days is on top of a 4% gelatin block. After isolation of the photoreceptor layer (200 µm) by vibratome, the photoreceptors are seeded after mechanical and enzymatic dissociation for culture. The photoreceptor layer can be used for molecular, biochemical analyses or transplantation.

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Immunology and Infection

Large-Scale Purification of Porcine or Bovine Photoreceptor Outer Segments for Phagocytosis Assays on Retinal Pigment Epithelial Cells
Célia Parinot 1,2,3, Quentin Rieu 1,2,3, Jonathan Chatagnon 1,2,3, Silvia C. Finnemann 4, Emeline F. Nandrot 1,2,3
1INSERM, U968, 2Sorbonne Universités, UPMC Paris 06, UMR_S 968, Institut de la Vision, 3CNRS, UMR_7210, 4Department of Biological Sciences, Center for Cancer, Genetic Diseases and Gene Regulation, Fordham University

This article describes the protocol for the purification of photoreceptor outer segment fragments (POS) via ultracentrifugation from porcine/bovine retinae using homogenization and sucrose gradient centrifugation. This protocol allows the preparation of large stocks of POS aliquots, labeled or unlabeled, that can then be stored at -80 °C.

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Medicine

Using Adeno-associated Virus as a Tool to Study Retinal Barriers in Disease
Ophélie Vacca 1,2,3, Brahim El Mathari 1,2,3, Marie Darche 1,2,3, José-Alain Sahel 1,2,3, Alvaro Rendon 1,2,3, Deniz Dalkara 1,2,3
1Department of Therapeutics, Institut de la Vision, Sorbonne Universtés, UPMC Univ Paris 06, UMR_S 968, 2INSERM, U968, 3CNRS, UMR_7210

To investigate the blood-retinal barrier permeability and the inner limiting membrane integrity in animal models of retinal disease, we used several adeno-associated virus (AAV) variants as tools to label retinal neurons and glia. Virus mediated reporter gene expression is then used as an indicator of retinal barrier permeability.

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Developmental Biology

Defined Xeno-free and Feeder-free Culture Conditions for the Generation of Human iPSC-derived Retinal Cell Models
Amélie Slembrouck-Brec *1, Céline Nanteau *1, José-Alain Sahel 1, Olivier Goureau 1, Sacha Reichman 1
1Institut de la Vision, Sorbonne Université, INSERM, CNRS, F-75012 Paris, France

The production of specialized retinal cells from pluripotent stem cells is a turning point in the development of stem cell-based therapy for retinal diseases. The present paper describes a simple method for an efficient generation of retinal organoids and retinal pigmented epithelium for basic, translational, and clinical research.

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Neuroscience

Cone-Enriched Cultures from the Retina of Chicken Embryos to Study Rod to Cone Cellular Interactions
Géraldine Millet-Puel 1, Myriam Pinault 1, Marie Cordonnier 1, Valérie Fontaine 1, José-Alain Sahel 1, Thierry Léveillard 1
1Department of Genetics - Sorbonne Université, INSERM, CNRS, Institut de la Vision

We describe a method to obtain primary cultures of cone photoreceptors from the retina of chicken embryos and its use for high content screening.

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