Three-Dimensional Culture Decreases the Angiogenic Ability of Mouse Macrophages
Haoxin Shi1, Dong Li2, Qing Shi2
1Endoscopy Room, Department of Gastroenterology, Shandong Cancer Hospital and Institute, Shandong First Medical University and Shandong Academy of Medical Science, Jinan, China.
Abstract:
Macrophages play important roles in angiogenesis; however, previous studies on macrophage angiogenesis have focused on traditional 2D cultures. In this study, we established a 3D culture system for macrophages using collagen microcarriers and assessed the effect of 3D culture on their angiogenic capabilities. Macrophages grown in 3D culture displayed a significantly different morphology and arrangement under electron microscopy compared to those grown in 2D culture. Tube formation assays and chick embryo chorioallantoic membrane assays further revealed that 3D-cultured macrophages were less angiogenic than those in 2D culture. Whole-transcriptome sequencing showed that nearly 40% of genes were significantly differently expressed, including nine important angiogenic factors of which seven had been downregulated. In addition, the expression of almost all genes related to two important angiogenic pathways was decreased in 3D-cultured macrophages, including the two key angiogenic factors, VEGFA and ANG2. Together, the findings of our study improve our understanding of angiogenesis and 3D macrophage culture in tissues, and provide new avenues and methods for future research on macrophages.
Insights
Three-dimensional (3D) macrophage culture using collagen microcarriers significantly alters cell morphology and reduces angiogenic potential compared to traditional 2D methods. This 3D system offers new insights into macrophage behavior in tissue-like environments.
Area of Science:
- Cell Biology
- Biotechnology
- Tissue Engineering
Background:
- Macrophages are crucial for angiogenesis, the formation of new blood vessels.
- Previous research on macrophage angiogenesis primarily utilized 2D cell culture models.
- The limitations of 2D cultures in mimicking in vivo tissue environments are well-documented.
Purpose of the Study:
- To establish and evaluate a 3D macrophage culture system using collagen microcarriers.
- To investigate the impact of 3D culture on macrophage morphology and angiogenic capabilities.
- To compare the angiogenic potential of macrophages cultured in 3D versus 2D environments.
Main Methods:
- Development of a 3D macrophage culture system utilizing collagen microcarriers.
- Morphological and ultrastructural analysis of macrophages using electron microscopy.
- Assessment of angiogenic potential via in vitro tube formation assays and in vivo chick embryo chorioallantoic membrane assays.
- Whole-transcriptome sequencing to identify differential gene expression.
Main Results:
- Macrophages in 3D culture exhibited distinct morphology and arrangement compared to 2D cultures.
- 3D-cultured macrophages demonstrated significantly reduced angiogenic capacity in both tube formation and chorioallantoic membrane assays.
- Whole-transcriptome analysis revealed differential expression in nearly 40% of genes, including downregulation of key angiogenic factors like VEGFA and ANG2.
- Expression of genes within major angiogenic pathways was notably decreased in 3D-cultured macrophages.
Conclusions:
- The 3D macrophage culture system provides a more physiologically relevant model for studying macrophage-mediated angiogenesis.
- Macrophages cultured in 3D exhibit suppressed angiogenic potential compared to those in 2D culture.
- These findings enhance understanding of macrophage behavior in tissue contexts and offer novel research methodologies.
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