Related Experiment Video
Updated: Jan 19, 2026
Rapid Isolation of Dorsal Root Ganglion Macrophages
Published on: September 7, 2019
Rapid Isolation of Dorsal Root Ganglion Macrophages
Xiaobing Yu1, Jacqueline Leff2, Zhonghui Guan3
1Department of Anesthesia and Perioperative Care, University of California San Francisco; xiaobing.yu@ucsf.edu.
Abstract:
There are growing interests to study the molecular and cellular interactions among immune cells and sensory neurons in the dorsal root ganglia after peripheral nerve injury. Peripheral monocytic cells, including macrophages, are known to respond to a tissue injury through phagocytosis, antigen presentation, and cytokine release. Emerging evidence has implicated the contribution of dorsal root ganglia macrophages to neuropathic pain development and axonal repair in the context of nerve injury. Rapidly phenotyping (or "rapid isolation of") the response of dorsal root ganglia macrophages in the context of nerve injury is desired to identify the unknown neuroimmune factors. Here we demonstrate how our lab rapidly and effectively isolates macrophages from the dorsal root ganglia using an enzyme-free mechanical dissociation protocol. The samples are kept on ice throughout to limit cellular stress. This protocol is far less time consuming compared to the standard enzymatic protocol and has been routinely used for our Fluorescence-activated Cell Sorting analysis.
Insights
Researchers developed a fast, enzyme-free method to isolate dorsal root ganglia macrophages after nerve injury. This technique aids in studying neuroimmune factors contributing to neuropathic pain and nerve repair.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Peripheral nerve injury triggers complex neuroimmune responses in the dorsal root ganglia (DRG).
- Macrophages within the DRG play a critical role in neuropathic pain development and axonal regeneration.
- Efficient isolation of DRG macrophages is crucial for understanding these neuroimmune interactions.
Purpose of the Study:
- To develop and validate a rapid, enzyme-free protocol for isolating macrophages from the dorsal root ganglia.
- To facilitate timely phenotyping of DRG macrophages for identifying novel neuroimmune factors.
Main Methods:
- Enzyme-free mechanical dissociation of DRG tissue.
- Maintaining samples on ice to minimize cellular stress.
- Application of the isolated cells in Fluorescence-activated Cell Sorting (FACS) analysis.
Main Results:
- Successful isolation of viable DRG macrophages using the mechanical protocol.
- The protocol is significantly faster than traditional enzymatic methods.
- The method is robust and suitable for downstream molecular analyses like FACS.
Conclusions:
- The developed enzyme-free mechanical dissociation protocol provides a rapid and effective means to isolate DRG macrophages.
- This method enhances the study of neuroimmune responses in neuropathic pain and nerve repair.
- Facilitates rapid phenotyping for identifying key factors in nerve injury.
