Protocol to develop a 3D mouse autologous lung tumor-immune spheroid co-culture model to advance radiotherapy-based treatments

Research Overview

3D culture reproduces tumor structure and cell interactions far better than monolayers, which matters for therapy research. This protocol builds a 3D mouse autologous lung tumor–immune spheroid co-culture to advance radiotherapy-based treatment.

It covers optimizing tumor spheroid formation in agarose molds, isolating CD4+ T cells and monocytes, differentiating regulatory T cells and M2-like macrophages, seeding the immune cells into tumor spheroids, characterizing the resulting tumor–immune spheroids with cell trackers and flow cytometry, and finally assessing radiotherapy efficacy in the co-culture.