Lowy Medical Research Institute

4 ARTICLES PUBLISHED IN JoVE

Developmental Biology

Efficient Derivation of Retinal Pigment Epithelium Cells from Stem Cells

Peter Westenskow^1,2, Zack Sedillo^1,2, Ashley Barnett², 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.

Medicine

Performing Subretinal Injections in Rodents to Deliver Retinal Pigment Epithelium Cells in Suspension

Peter D. Westenskow^1,2, Toshihide Kurihara¹, Stephen Bravo¹, Daniel Feitelberg¹, Zack A. Sedillo², Edith Aguilar¹, 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.

Immunology and Infection

Assessing Retinal Microglial Phagocytic Function In Vivo Using a Flow Cytometry-based Assay

Salome Murinello¹, Stacey K. Moreno¹, Matthew S. Macauley², Susumu Sakimoto¹, Peter D. Westenskow^1,3, Martin Friedlander¹

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.

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).