Abstract
Background Recent studies have shown the obvious interaction between cancer cells and immune cells. These effects can be hardly studied in conventional culture systems using cell monolayers. Therefore, alternative in vitro models are required. We describe a novel 3D cell culture model and a 3D migration assay for investigating cancer cell/immune cell interactions. Methods A multi-well hanging drops system was used to produce 3D tumour spheres. The human NSCLC cell lines A549, Calu-6 and Colo699 were incubated for 6 and 7 days in the hanging drops to form spheroids. On day 5, peripheral blood mononuclear cells (PBMC) were added either with or without interleukin-2 (IL-2). Viability was investigated via flowing cytometry. Invasion of PBMC into the tumour spheres was measured by performing immunohistochemistry. Results No effect on cell viability was observed in A549 and Colo699 spheroids. In Calu-6 spheroids, a significant PBMC induced cytotoxic effect was measured, what was even stronger under IL-2 stimulation. Immunohistochemical staining revealed PBMC infiltration in all three cell lines. Under IL-2 stimulation, infiltration of PBMC increased in Calu-6 and Colo699 spheroids. Conclusion This work provides evidence that our 3D co-culture model is a reliable and effective approach to study interactions between cancer cells and immune cells. Additionally, we were able to establish for the first time an innovative ex vivo infiltration assay based on 3D microtissues.
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