Researchers from the Germans Trias i Pujol Research Institute (IGTP) have published a study in Scientific Reports (Nature Portfolio) that provides an analysis of three-dimensional (3D) culture techniques for generating multicellular tumour 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 tumour behaviour and drug response under conditions that more closely resemble real tumour 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 standardise 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 tumour-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 optimisation and systematic comparison of 3D culture techniques. We believe it can serve as a guide for other groups working with tumour models in vitro and help standardise protocols that facilitate more robust and comparable studies in preclinical colorectal cancer research, " highlights Sergio Alonso, the last author and corresponding author of the study.
Scientific Reports
Experimental study
Cells
Comparative analysis of 3D-culture techniques for multicellular colorectal tumour spheroids and development of a novel SW48 3D-model
29-Jul-2025
The authors declare no competing interests.