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JoVE Journal
Biology
Primary Cell Cultures from the Mouse Retinal Pigment Epithelium
Primary Cell Cultures from the Mouse Retinal Pigment Epithelium
JoVE Journal
Biology
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JoVE Journal Biology
Primary Cell Cultures from the Mouse Retinal Pigment Epithelium

Primary Cell Cultures from the Mouse Retinal Pigment Epithelium

Full Text
22,323 Views
09:35 min
March 16, 2018

DOI: 10.3791/56997-v

Peng Shang1, Nadezda A. Stepicheva1, Stacey Hose1, J. Samuel Zigler, Jr.2, Debasish Sinha1,2

1Department of Ophthalmology,University of Pittsburgh School of Medicine, 2Wilmer Eye Institute,The Johns Hopkins University School of Medicine

The retinal pigment epithelium (RPE) is a multi-functional epithelium of the eye. Here we present a protocol to establish primary cell cultures derived from the murine RPE.

The overall goal of this procedure is to demonstrate a feasible way to obtain retinal pigment epithelium cells from mouse eyes, and to culture them in vitro while retaining their original properties. This method will help RPE researchers to obtain dissection skills that are required for establishing of mouse RPE cell cultures, and to demonstrate how to do that in an easy way. The main advantage of this procedure is that it is simple and feasible.

The only thing required is practice, and the visual demonstration of RPE isolation and the microscopy literally demonstrates how to obtain the required skills. To isolate the mouse eyeballs, first euthanize two mice using any approved method, and wearing gloves place it on an absorbent pad. Position a thumb and index finger around the eye and gently push the skin to bulge out the eyeball.

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