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Lowy Medical Research Institute

4 ARTICLES PUBLISHED IN JoVE

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Developmental Biology

Efficient Derivation of Retinal Pigment Epithelium Cells from Stem Cells
Peter Westenskow 1,2, Zack Sedillo 1,2, Ashley Barnett 2, Martin Friedlander 1,2
1Department of Cell and Molecular Biology, The Scripps Research Institute, 2Lowy Medical Research Institute

Stem cell-derived retinal pigment epithelium (RPE) cells may be used for multiple applications including cell-based therapies for retinal degeneration, disease modeling, and drug studies. Here we present a simple protocol for reproducibly deriving RPE from stem cells.

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Medicine

Performing Subretinal Injections in Rodents to Deliver Retinal Pigment Epithelium Cells in Suspension
Peter D. Westenskow 1,2, Toshihide Kurihara 1, Stephen Bravo 1, Daniel Feitelberg 1, Zack A. Sedillo 2, Edith Aguilar 1, Martin Friedlander 1,2
1Department of Cell and Molecular Biology, The Scripps Research Institute, 2Lowy Medical Research Institute

Here we present a community accepted protocol in multimedia format for subretinally injecting a bolus of RPE cells in rats and mice. This approach can be used for determining rescue potentials, safety profiles, and survival capacities of grafted RPE cells upon implantation in animal models of retinal degeneration.

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Immunology and Infection

Assessing Retinal Microglial Phagocytic Function In Vivo Using a Flow Cytometry-based Assay
Salome Murinello 1, Stacey K. Moreno 1, Matthew S. Macauley 2, Susumu Sakimoto 1, Peter D. Westenskow 1,3, Martin Friedlander 1
1Department of Cell and Molecular Biology, The Scripps Research Institute, 2Department of Chemical Physiology, The Scripps Research Institute, 3The Lowy Medical Research Institute

Microglial phagocytosis is critical for the maintenance of tissue homeostasis and inadequate phagocytic function has been implicated in pathology. However, assessing microglia function in vivo is technically challenging. We have developed a simple but robust technique for precisely monitoring and quantifying the phagocytic potential of microglia in a physiological setting.

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Bioengineering

Toxicity Screens in Human Retinal Organoids for Pharmaceutical Discovery
Kevin Eade 1,2, Sarah Giles 1,2, Sarah Harkins-Perry 1,2, Martin Friedlander 1,2
1Lowy Medical Research Institute, 2The Scripps Research Institute

Here we present a step-by-step protocol to generate mature human retinal organoids and utilize them in a photoreceptor toxicity assay to identify pharmaceutical candidates for the age-related retinal degenerative disease macular telangiectasia type 2 (MacTel).

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