HMN 2025: How Colorectal cancer study expands and optimizes 3D tumor models

Researchers from the Germans Trias i Pujol Research Institute (IGTP) have published a study in Scientific Reports that provides an analysis of three-dimensional (3D) culture techniques for generating multicellular tumor spheroids (MCTS) of colorectal cancer. The study, authored entirely by members of the Cancer Genetics and Epigenetics (CGE) group, lists Dr. Sergio Alonso and Dr. Beatriz González as corresponding authors.

3D models allow for a more accurate study of tumor behavior and drug response under conditions that more closely resemble real tumor tissue. The team compared three common methods (liquid overlay, hanging drop, and U-bottom plates) across eight colorectal cancer cell lines, using different matrices (Matrigel, collagen I, and methylcellulose).

The results show that the morphology and compactness of the spheroids depend both on the cell line and the culture conditions, and highlight the need to standardize protocols. The study describes, for the first time, a compact model using the SW48 cell line, which so far had been considered non-spheroid-forming. In addition, it demonstrates that treating standard plates with an anti-adherent solution enables the generation of spheroids in a more cost-effective and scalable manner than using commercially available cell-repellent plates.

It also highlights the incorporation of colonic fibroblasts in co-culture, which increases the biological relevance of the models and allows better simulation of tumor-stroma interactions. The analysis has also revealed notable morphological differences between isogenic lines (DLD1/HCT8 and SW480/SW620), which points to transcriptional or epigenetic factors in the formation of spheroids.

“This work is the result of many months of optimization and systematic comparison of 3D culture techniques. We believe it can serve as a guide for other groups working with tumor models in vitro and help standardize protocols that facilitate more robust and comparable studies in preclinical colorectal cancer research,” says Alonso, the last author and corresponding author of the study.