Table 1.

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

Co‐culture settings
Matrix	Device (methods)	Cell types	Application	Technology	Ref
DOUBLE CULTURES, Cancer cells and fibroblasts
Collagen I	Transwell with mouse intestinal basal membrane. Lower compartment, fibroblast mixed in collagen; upper compartment, tumor cells	HT29, HCT116; patient derived CAFs and NFs	Invasion studies	IF; microscopy methods; time‐lapse imaging	[155]
BME; Matrigel	Transwell. Lower compartment, fibroblasts seeded directly or embedded in BME; upper compartment, organoids in matrigel droplets	Mouse and human CRC organoids; primary mouse colon fibroblasts	Tumor‐stroma interaction studies	qRT‐PCR analysis; contraction assay	[156]
Matrigel	Transwell. Lower compartment, fibroblasts; upper compartment, organoids in matrigel droplets	PDO and CAFs from surgical specimens of CRCs	Gene expression profiles	DNA microarray; viability assays	[165]
Collagen	Tumoroids in two layers of collagen gel	DLD‐1; human primary lung cancer‐derived fibroblasts	Invasion studies	Fluorescence imaging; IHC; time‐lapse experiments	[150]
	Tumoroids formed in agarose coated plates	SW480; MRC5	Drug response analysis; secretomes analysis	Fluorescence imaging; LC‐MS; viability assays	[157]
	Tumoroids formed in polydimethylsiloxane (PDMS) microwell platform.	HCT‐8; NIH3T3	Drug response analysis; co‐culture self‐organization; stiffness variation	Immunostaining; time‐laps imaging; mechanical stability measurement; WB analysis	[158]
	Tumoroids formed in hanging drop plates	HCT116; CCD‐18Co	High‐content drug discovery	Genomics screening; Imaging analysis; RNAseq	[168]
BME; Matrigel	Matrix droplet, Tumor cells and CAFs plated together. Transwell. Lower compartment, fibroblasts embedded in BME; upper compartment, organoids in matrigel droplets	PDO and CAFs from surgical specimens of CRCs	Tumor‐stroma interaction studies; drug response analysis	qRT‐PCR analysis; RNAseq; viability assays;	[163]
Collagen I	Matrix droplet into a force sensor	FET; human primary colorectal CAFs	Cell forces and tissue stiffness measurement	High‐resolution force sensor	[154]
Matrigel; Collagen I	Organoids plated in Matrix droplet and CAFs in suspension in the well	PDO and CAFs from surgical specimens of CRC liver metastasis	Tumor‐stroma interaction studies; mesenchymal‐like cancer model	scRNAseq; IF; live cell imaging; cell culture medium assays	[161]
hyaluronan‐gelatin hydrogels	CAFs seeded on top of each PDO Matrix droplet	PDO and CAFs from surgical specimens of CRCs	Drug response analysis; gene expression profiles	RNAseq; viability assays; immunostaining	[164]
Matrigel; Collagen I	Tumor cells and fibroblasts plated together in Matrix droplet or divided into individual layers in ALI culture.	PDO, NFs and CAFs from surgical specimens of CRCs	Tumor‐stroma interaction studies	Immunostaining; RNAseq; Proteomic; 3D migration assay; Cytokine array	[162]
GelMA	Bioprinting, PDO and CAF microtissue formed by acoustic droplets technology	PDO and CAFs from surgical specimens of CRCs	Migration and invasion models; drug response analysis	Time‐lapse imaging; IF	[167]