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Updated: May 7, 2026
Isolation of Cortical Microglia with Preserved Immunophenotype and Functionality From Murine Neonates
Published on: January 30, 2014
Human amnion mesenchymal cells negative co-stimulatory molecules PD-L1 expression and its capacity of modulating
Weijiang Wu1, Qing Lan, Hua Lu
1Department of Neurosurgery, The Second Affiliated Hospital of Soochow University, Suzhou, 215004, China, wuwj2004@sohu.com.
Abstract:
The objective of this study is to investigate the negative immunomodulatory capacity of human amniotic mesenchymal cells (AMSCs) and their possible intrinsic mechanism, by which we can confirm that they modulate microglial activation of central nervous system from multiple perspectives at the molecular level. The identification of the immune phenotype of AMSCs and microglial cells was executed by immunohistochemical methods and flow cytometry. Meanwhile, the influence and mechanism of amniotic mesenchymal cells in vitro on proliferation, cell cycle, and cytokine release of activated microglia (MI) would be detected by ELISA, β-liquid scintillation counting method, and flow cytometry. Human amnion mesenchymal cells highly expressed negative co-stimulatory molecules PD-L1, while its ligand PD1 was expressed with high level by activated MI. When adding the PD-L1mAb to the mixed culture system composed of AMSCs and activated MI, the proliferation inhibitory effect and the cycle-blocking effect produced by the former on the latter would be partially reversed; at the same time, the impact of the latter cytokine secretion would be adjusted. As a conclusion, AMSCs play inhibitory effects on microglial activation, proliferation, and immune effects partially through the PD-L1-PD1 signaling pathways.
Insights
Human amniotic mesenchymal stem cells (AMSCs) inhibit central nervous system microglial activation. This immunomodulatory effect is partly mediated by the PD-L1/PD-1 pathway, impacting microglial proliferation and cytokine release.
Area of Science:
- Immunology
- Neuroscience
- Stem Cell Biology
Background:
- Microglia play a crucial role in central nervous system immunity.
- Dysregulated microglial activation contributes to neuroinflammation.
- Human amniotic mesenchymal stem cells (AMSCs) possess immunomodulatory properties.
Purpose of the Study:
- To investigate the negative immunomodulatory capacity of AMSCs on microglial activation.
- To elucidate the molecular mechanisms underlying AMSC-mediated modulation of microglia.
- To confirm the effect of AMSCs on central nervous system immune cells.
Main Methods:
- Immunohistochemistry and flow cytometry for immune phenotyping of AMSCs and microglia.
- ELISA, beta-liquid scintillation counting, and flow cytometry to assess microglial proliferation, cell cycle, and cytokine release in vitro.
- Involvement of PD-L1/PD-1 signaling pathway investigated using PD-L1 monoclonal antibody (mAb).
Main Results:
- AMSCs highly express programmed death-ligand 1 (PD-L1), while activated microglia express its receptor, programmed death 1 (PD-1).
- AMSCs exhibit inhibitory effects on microglial proliferation and cell cycle progression.
- Blocking the PD-L1/PD-1 interaction partially reversed the inhibitory effects of AMSCs on microglia and modulated cytokine secretion.
Conclusions:
- AMSCs exert inhibitory effects on microglial activation, proliferation, and immune responses.
- The PD-L1/PD-1 signaling pathway is a key mechanism mediating the immunomodulatory effects of AMSCs on microglia.
- AMSCs hold potential for therapeutic applications in neuroinflammatory conditions.
