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Updated: Feb 25, 2026
Investigation of Macrophage Polarization Using Bone Marrow Derived Macrophages
Published on: June 23, 2013
Differently polarized macrophages affect the viability and growth of NSPCs by regulating the expression of PACAP
Min Liu1, Ming Deng2, Yonggang Ma2
1Department of Immunology, School of Medicine, Wuhan University, Wuhan 430071, China.
Abstract:
Objective To explore the influence of differently polarized macrophages, M1 or M2, to viability and growth of NSPCs and its possible mechanism, especially the role of pituitary adenylate cyclase-activating polypeptide (PACAP) in it.
Method:
Spinal cord-derived NSPCs were co-cultured with M1 or M2 through a transwell system. Growth of NSPCs in both groups was observed through an inverted microscope within 3days. NSPCs viability of each group, represented as intracellular ATP levels, was measured using the Cellular ATP Kit HTS following co-culture for 24h. PACAP levels in the co-cultured NSPCs was alleviated with immunofluorescence and Western blot analysis.
Results:
Morphologically NSPCs demonstrated a long spindle shape with short sprout on 3rd day when cultured together with M2. NSPCs cultured with M1 showed a small circle or oval shape with no obvious sprout. Expression of PACAP was observed in NSPCs co-cultured with M2 through immunofluorescence. In contrast, NSPCs did not demonstrate PACAP staining in the presence of M1 or cultured alone. PACAP in the NSPCs co-cultured with M2 was upregulated significantly compared with that co-cultured with M1 according to Western blot method. The relative ATP level of NSPCs co-cultured with M1 was markedly decreased while that with M2 was elevated significantly. That trend could be relieved by exogenous PACAP or PACAP 6-38. Viability change of NSPCs by M1/M2 correlated with apoptosis.
Conclusion:
Differently polarized macrophages could affect the growth and viability of NSPCs by regulating the expression of PACAP.
Insights
Macrophages M1 and M2 polarization influence neural stem cell (NSPCs) viability and growth. This effect is mediated by regulating pituitary adenylate cyclase-activating polypeptide (PACAP) expression.
Area of Science:
- Neuroscience
- Immunology
- Stem Cell Biology
Background:
- Neural stem cells (NSPCs) are crucial for central nervous system repair.
- Macrophages play diverse roles in the central nervous system, including modulating NSPC behavior.
- Understanding macrophage polarization (M1 vs. M2) effects on NSPCs is vital for regenerative medicine.
Purpose of the Study:
- To investigate how M1 and M2 polarized macrophages impact NSPC viability and growth.
- To elucidate the underlying molecular mechanisms, focusing on pituitary adenylate cyclase-activating polypeptide (PACAP).
Main Methods:
- Spinal cord-derived NSPCs were co-cultured with M1 or M2 macrophages using a transwell system.
- NSPC growth was monitored via microscopy; viability was assessed by intracellular ATP levels.
- PACAP expression was analyzed using immunofluorescence and Western blot.
Main Results:
- M2 macrophages promoted NSPC growth and viability, indicated by spindle morphology and increased ATP levels.
- M1 macrophages impaired NSPC growth and viability, showing oval morphology and decreased ATP levels.
- PACAP expression was upregulated in NSPCs co-cultured with M2 macrophages, correlating with enhanced viability.
Conclusions:
- Differential macrophage polarization (M1/M2) significantly affects NSPC growth and viability.
- PACAP acts as a key mediator in the interaction between polarized macrophages and NSPCs.
- Targeting PACAP may offer therapeutic strategies for NSPC-based therapies.
