Table 2.

Schematic summary of technical approaches exploited for double cultures of CR‐cancer cells and immune cells.

Co‐culture settings
Matrix	Device (methods)	Cell types	Application	Technology	Ref
DOUBLE CULTURES, Cancer cells and immune cells
Geltrex	PBMC + single cell‐dissociated organoids (20, 1 E, T ratio)	PDO and PBMC from surgical specimens of CRCs	Cancer‐T cells interaction studies	Flow cytometry; cytotoxicity assays	[170, 171]
BME; Matrigel	T cells + HT29 (1, 10 ratio); T cells + PDO	PDO from surgical specimens of healty colon or tumoral tissue; healthy donor PBMC (T cells); HT29; Colo205; Jurkat	Cancer‐T cells interaction studies; immune surveillance	Time‐lapse imaging, immunofluorescence	[172]
Matrigel	CD8 T cells + tumor cells (2, 1 or 5, 1 E, T ratios)	PDO from surgical specimens of CRCs; healthy donor PBMC (CD8+ Tcells)	Immunotherapy response analysis	Fluorescence imaging; flow cytometry	[173]
	Vδ2 T cells + CRC spheroids (1, 1 E, T ratio).	HCT15, SW620, DLD1 spheroids; healthy donors PBMC (Vδ2 T cells)	Immunotherapy response analysis	Viability and cytotoxicity assays; spheroid size measurement	[174]
	CD19−CD14− sorted PBMCs or TILs + tumor cells (1, 1 E, T ratio)	HT‐29, DLD1; healthy donors CD19−CD14− sorted PBMCs; patient‐derived tumor spheroids; autologous tumor‐infiltrating lymphocytes (TILs)	Immune infiltrate characterization; antitumor effects studies	Flow cytometry; IF; spheroid volume calculation	[175]
	CRC spheroids + human monocytes (1, 1 E, T ratio).	SW620‐CSC; healthy donors PBMC (monocytes)	Analysis of tumor hybrid cells properties	Migration assay; viability assay; flow cytometry; soluble immune‐checkpoint measurement	[177]
Matrigel	Organoids seeded on a Matrigel layer or on a layer of primary human colon fibroblasts + NK‐92 cells	PDO from surgical specimens of CRCs; normal PDO and primary fibroblasts from non‐pathological mucosa	CAR‐NK immunotherapy studies	Fluorescence imaging; viability and cytotoxicity assays	[179]