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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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