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).