Isolation of Macrophages from Mouse Dorsal Root Ganglion Using Mechanical Dissociation

Published: July 31, 2023

Abstract

Source: Yu, X., et al. Rapid Isolation of Dorsal Root Ganglion Macrophages. J. Vis. Exp. (2019)

This video demonstrates an enzyme-free protocol to isolate macrophages from mouse dorsal root ganglion (DRG) using a mechanical dissociation technique. Upon homogenization of the freshly collected DRG tissue, antibodies against the cell-surface receptors on the macrophages are used to label the cells fluorescently, followed by performing flow cytometry to confirm the presence of macrophages in the sample.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. All animal experiments were approved by the Institutional Animal Care and Use Committee at the University of California San Francisco and were conducted in accordance with the NIH Guide for the Care and Use of Laboratory Animals. 1. Collect lumbar DRG from experimental mice Befo…

Divulgaciones

The authors have nothing to disclose.

Materials

AP20187 Clontech 635058
a-mouse CX3CR1-APC antibody Biolegend 149007
Avertin Sigma T48402
Cell strainer (70 mm nylon) Falcon 352350
Centrifuge Eppendorf 5810R
Dounce tissue homogenizer Wheaton 357538 (1ml)
FACS tubes (5ml) Falcon 352052
Friedman-Pearson Rongeur FST 16121-14
HBSS (10x, Ca++/Mg++-free) Gibco 14185-052
Noyes Spring Scissor FST 15012-12
Percoll Sigma P4937-500ml
Propidium iodide Sigma P4864-10ml

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Isolation of Macrophages from Mouse Dorsal Root Ganglion Using Mechanical Dissociation. J. Vis. Exp. (Pending Publication), e21517, doi: (2023).

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