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Selective depletion of cultured macrophages by magnetite nanoparticles modified with gelatin
Yoshihiro Komohara1, Ryuta Kawauchi2, Erika Makiyama2
1Department of Cell Pathology, Graduate School of Medical Sciences, Kumamoto University, Kumamoto 860-8556, Japan.
Abstract:
Previous studies have indicated pro-tumor functions of macrophages in tumor progression in different types of malignant tumors. The detailed mechanisms of cell-cell interaction between macrophages and tumor cells have been investigated by means of in vitro co-culture experiments. The present study developed magnetite nanoparticles modified with gelatin that are specifically engulfed by macrophages and investigated methods to deplete these macrophages in co-culture experiments using a magnet. T98G glioma cell line and human monocyte-derived macrophages were mixed and co-cultured for 2 days. The T98G cells were isolated by depletion of the macrophages using the magnetite nanoparticles. mRNA expression of a number of pro-tumor molecules in the isolated T98G cells, with or without co-culture with macrophages, was then evaluated. The mRNA expression levels of chemokine (CC motif) ligand 2, interleukin-6 and macrophage-colony stimulating factor receptor (M-CSFR) were significantly upregulated in T98G cells by co-culture with macrophages (P<0.01). M-CSFR protein expression was also increased by co-culture with macrophages. The conditioned medium of co-cultured cells increased M-CSFR expression in T98G cells. Magnetite nanoparticles may be a novel tool not only for investigating the unique activation status of tumor cells in co-culture conditions, but also for targeting pro-tumor macrophages in tumor tissues.
Insights
Magnetite nanoparticles help isolate tumor cells by removing pro-tumor macrophages in co-culture. This method reveals how macrophages enhance glioma cell expression of key molecules like interleukin-6.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Macrophages play a significant role in promoting tumor progression across various cancers.
- Understanding macrophage-tumor cell interactions is crucial for developing effective cancer therapies.
Purpose of the Study:
- To develop and evaluate gelatin-modified magnetite nanoparticles for macrophage depletion in co-culture systems.
- To investigate the impact of macrophage co-culture on glioma (T98G) cell gene expression.
- To assess the potential of magnetite nanoparticles as a tool for studying tumor microenvironments.
Main Methods:
- Co-culture of T98G glioma cells with human monocyte-derived macrophages.
- Depletion of macrophages using gelatin-modified magnetite nanoparticles and magnetic separation.
- Quantitative mRNA expression analysis of pro-tumor molecules (CCL2, IL-6, M-CSFR) in isolated T98G cells.
- Assessment of M-CSFR protein expression and effect of conditioned medium.
Main Results:
- Co-culture with macrophages significantly upregulated mRNA expression of chemokine (CC motif) ligand 2, interleukin-6, and macrophage-colony stimulating factor receptor (M-CSFR) in T98G cells (P<0.01).
- Macrophage co-culture also increased M-CSFR protein expression in T98G cells.
- Conditioned medium from co-cultured cells demonstrated an ability to increase M-CSFR expression in T98G cells.
Conclusions:
- Gelatin-modified magnetite nanoparticles provide a novel method for depleting macrophages in co-culture, enabling the study of tumor cell activation.
- Macrophages significantly enhance the expression of pro-tumor molecules in T98G glioma cells.
- Magnetite nanoparticles show promise for both research applications and potential therapeutic targeting of pro-tumor macrophages in vivo.
