Selective depletion of cultured macrophages by magnetite nanoparticles modified with gelatin

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.