Isolation of Primary Murine Retinal Ganglion Cells (RGCs) by Flow Cytometry

Overview

Journal J Vis Exp

Specialties Biology
General Medicine

Date 2017 Jul 18

PMID 28715391

Citations 6

Authors

Sumana R Chintalapudi

Need N Patel

Zachary K Goldsmith

Levon Djenderedjian

Xiang Di Wang

Tony N Marion

Monica M Jablonski

Vanessa M Morales-Tirado

Affiliations

Soon will be listed here.

Abstract

Neurodegenerative diseases often have a devastating impact on those affected. Retinal ganglion cell (RGC) loss is implicated in an array of diseases, including diabetic retinopathy and glaucoma, in addition to normal aging. Despite their importance, RGCs have been extremely difficult to study until now due in part to the fact that they comprise only a small percentage of the wide variety of cells in the retina. In addition, current isolation methods use intracellular markers to identify RGCs, which produce non-viable cells. These techniques also involve lengthy isolation protocols, so there is a lack of practical, standardized, and dependable methods to obtain and isolate RGCs. This work describes an efficient, comprehensive, and reliable method to isolate primary RGCs from mice retinae using a protocol based on both positive and negative selection criteria. The presented methods allow for the future study of RGCs, with the goal of better understanding the major decline in visual acuity that results from the loss of functional RGCs in neurodegenerative diseases.

Citing Articles

Purification of retinal ganglion cells using low-pressure flow cytometry.

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Selective deletion of zinc transporter 3 in amacrine cells promotes retinal ganglion cell survival and optic nerve regeneration after injury.

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Selective retinal ganglion cell loss and optic neuropathy in a humanized mouse model of familial dysautonomia.

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Altered Energy Metabolism During Early Optic Nerve Crush Injury: Implications of Warburg-Like Aerobic Glycolysis in Facilitating Retinal Ganglion Cell Survival.

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