Related Experiment Video
Updated: Jul 4, 2026
Characterization and Isolation of Mouse Primary Microglia by Density Gradient Centrifugation
Published on: February 16, 2018
Repopulating Microglia Suppress Peripheral Immune Cell Infiltration to Promote Poststroke Recovery
Ligen Shi1,2,3, Lingxiao Lu1,3, Jun Hu1
1Department of Neurosurgery, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China.
Aims:
Sustained neuroinflammation following ischemic stroke impedes post-injury tissue repairment and neurological functional recovery. Developing innovative therapeutic strategies that simultaneously suppress detrimental inflammatory cascades and facilitate neurorestorative processes is critical for improving long-term rehabilitation outcomes.
Methods:
We employed a microglia depletion-repopulation paradigm by administering PLX5622 for 7 days post-ischemia; followed by a 7-day withdrawal period to allow microglia repopulation. Single-cell transcriptomics, behavioral testing, cytokine arrays, flow cytometry, and immunofluorescence were used to assess the effects of microglia repopulation and delineate the transition of reshaped immune microenvironment.
Results:
PLX5622 administration reshaped the poststroke immune microenvironment, promoting neurofunctional recovery. Repopulated microglia adopted a homeostatic phenotype, increasing homeostatic states by ~14.36% and reducing pro-inflammatory states by ~20.17%. This reshaped environment suppressed T cell exhaustion, limited neutrophil terminal differentiation, and promoted a phagocytic macrophage phenotype. Furthermore, we identified that these transitions in infiltrating immune cells may be driven by reduced chemokine production, enhanced blood-brain barrier (BBB) integrity, and transcriptional reprogramming.
Conclusion:
Transient microglial depletion and repopulation via PLX5622 during the acute phase post stroke facilitate the recovery of neurological function. This immunomodulatory strategy offers a promising and clinically translationally relevant approach to enhance functional recovery following ischemic brain injury.
Insights
Transiently depleting and repopulating microglia after ischemic stroke reshapes the immune environment, promoting neurofunctional recovery. This approach enhances brain repair and functional outcomes post-stroke.
Area of Science:
- Neuroscience
- Immunology
- Regenerative Medicine
Background:
- Sustained neuroinflammation after ischemic stroke hinders tissue repair and functional recovery.
- Novel therapeutic strategies are needed to suppress inflammation and promote neurorestoration.
Purpose of the Study:
- To investigate the effects of transient microglia depletion and repopulation on the post-stroke immune microenvironment.
- To assess the impact of this immunomodulatory strategy on neurological functional recovery.
Main Methods:
- Utilized a microglia depletion-repopulation paradigm using PLX5622 in a post-ischemic stroke model.
- Employed single-cell transcriptomics, behavioral testing, cytokine arrays, flow cytometry, and immunofluorescence.
Main Results:
- PLX5622 administration promoted neurofunctional recovery by reshaping the immune microenvironment.
- Repopulated microglia exhibited a more homeostatic phenotype, reducing pro-inflammatory states.
- Observed suppressed T cell exhaustion, limited neutrophil differentiation, and promoted phagocytic macrophages, potentially driven by reduced chemokine production and enhanced blood-brain barrier integrity.
Conclusions:
- Transient microglial depletion and repopulation facilitate neurological function recovery post-stroke.
- This immunomodulatory strategy shows promise for enhancing functional recovery in ischemic brain injury.
- The approach is clinically translatable for improving rehabilitation outcomes.
