JoVE Logo

Sign In

A subscription to JoVE is required to view this content. Sign in or start your free trial.

Abstract

Neuroscience

Enrichment of Adult Mouse Dorsal Root Ganglia Neuron Cultures by Immunopanning

Published: February 24th, 2023

DOI:

10.3791/64603

1Neuroscience and Mental Health Institute, University of Alberta, 2Department of Medicine, Division of Neurology, University of Alberta, 3Department of Medical Microbiology and Immunology, University of Alberta, 4Department of Pharmacology, University of Alberta, 5Department of Anesthesiology and Pain Medicine, University of Alberta

Abstract

Dorsal root ganglia (DRGs) are peripheral structures adjacent to the dorsal horn of the spinal cord, which house the cell bodies of sensory neurons as well as various other cell types. Published culture protocols often refer to whole dissociated DRG cultures as being neuronal, despite the presence of fibroblasts, Schwann cells, macrophages, and lymphocytes. While these whole DRG cultures are sufficient for imaging applications where neurons can be discerned based on morphology or staining, protein or RNA homogenates collected from these cultures are not primarily neuronal in origin. Here, we describe an immunopanning sequence for cultured mouse DRGs. The goal of this method is to enrich DRG cultures for neurons by removing other cell types. Immunopanning refers to a method of removing cell types by adhering antibodies to cell culture dishes. Using these dishes, we can negatively select against and reduce the number of fibroblasts, immune cells, and Schwann cells in culture. This method allows us to increase the percentage of neurons in cultures.

Explore More Videos

Keywords DRG Culture

This article has been published

Video Coming Soon

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved