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Updated: Dec 30, 2025
Mesenchymal Stem Cell Regulation of Macrophage Phagocytosis; Quantitation and Imaging
Published on: July 16, 2021
Human umbilical cord mesenchymal stem cells polarize RAW264.7 macrophages to an anti-inflammatory subpopulation
Yu Xu1, Hongxing Luo1, Fang Chen1
1Department of Cardiology, Zhengzhou University People's Hospital Zhengzhou, Henan, China.
Objective:
To investigate the effects of human umbilical cord mesenchymal stem cells (hUCMSCs) on the polarization of lipopolysaccharide-stimulated RAW264.7 macrophages.
Methods:
Lipopolysaccharide-stimulated RAW264.7 macrophages were co-cultured with hUCMSCs in a Transwell system for 4 d, and then labelled with anti-F4/80, anti-CD86, and anti-CD206 antibodies for flow cytometry. The co-cultured supernatants were detected by enzyme-linked immunosorbent assay for prostaglandin E2. The co-cultured RAW264.7 macrophages were also lysed to measure the intracellular level of inducible nitric oxide synthase.
Results:
There were significantly more F4/80+CD86+CD206+ RAW264.7 macrophages in the hUCMSCs-treated groups than the control group (P<0.001). The secretion of prostaglandin E2 by lipopolysaccharide-stimulated RAW264.7 macrophages was significantly inhibited in a dose-dependent manner with the addition of hUCMSCs (P<0.001). The expression of iNOS, the intracellular marker of M1 cells, was also significantly inhibited by hUCMSCs (P<0.05).
Conclusion:
hUCMSCs significantly polarize the lipopolysaccharide-stimulated RAW264.7 macrophages from a pro-inflammatory M1 subpopulation to an intermediate subpopulation of anti-inflammatory M2 macrophages, which are associated with a gradual decrease of iNOS and PGE2 levels.
Insights
Human umbilical cord mesenchymal stem cells (hUCMSCs) shift pro-inflammatory macrophages towards an anti-inflammatory state. This study shows hUCMSCs reduce inflammatory markers like iNOS and PGE2 in stimulated macrophages.
Area of Science:
- Immunology
- Stem Cell Biology
Background:
- Macrophages play a critical role in immune responses.
- Lipopolysaccharide (LPS) stimulation induces pro-inflammatory M1 macrophage polarization.
- Mesenchymal stem cells (MSCs) possess immunomodulatory properties.
Purpose of the Study:
- To investigate the immunomodulatory effects of human umbilical cord mesenchymal stem cells (hUCMSCs) on lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages.
- To determine if hUCMSCs can alter macrophage polarization.
- To assess the impact of hUCMSCs on inflammatory mediator production.
Main Methods:
- RAW264.7 macrophages were stimulated with LPS and co-cultured with hUCMSCs using a Transwell system.
- Flow cytometry was employed to analyze macrophage surface markers (F4/80, CD86, CD206).
- Enzyme-linked immunosorbent assay (ELISA) measured prostaglandin E2 (PGE2) levels, and inducible nitric oxide synthase (iNOS) expression was quantified.
Main Results:
- hUCMSCs significantly increased the population of F4/80+CD86+CD206+ macrophages.
- hUCMSCs dose-dependently inhibited PGE2 secretion from LPS-stimulated macrophages.
- hUCMSCs significantly reduced the expression of iNOS, a marker of M1 macrophages.
Conclusions:
- hUCMSCs effectively polarize LPS-stimulated RAW264.7 macrophages towards an intermediate M2-like phenotype.
- This polarization is associated with decreased levels of pro-inflammatory mediators, iNOS and PGE2.
- hUCMSCs exhibit potential as a therapeutic agent for modulating inflammatory responses.
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