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Significance Therapeutic strategies targeting glioblastoma (GBM) tumor-associated macrophages (TAMs) have shown limited efficacy. One reason for this limited effectiveness is our poor understanding of the fundamental characteristics driving the behavior of these cells. Here we report a combination of immunocompetent lineage tracing mouse models of GBM with an open skull window for in vivo observation of TAM constituents via 2-photon microscopy. We show that TAMs are composed of 2 morphologically and behaviorally distinct cell types: brain-resident microglia and bone marrow-derived macrophages (BMDMs). After treatment with anti-vascular endothelial growth factor A antibody, we observed significant reductions in BMDM infiltration and morphological responses. These studies provide a platform for monitoring the multidimensional responses of specific cell populations to therapeutic intervention in real time.

Intravital 2-photon imaging reveals distinct morphology and infiltrative properties of glioblastoma-associated macrophages