Isolation of Macrophages from Mouse Dorsal Root Ganglion Using Mechanical Dissociation

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…

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