Overview

• Researchers compared three 3D culture techniques using Matrigel, collagen I and methylcellulose across eight colorectal cancer cell lines to determine protocol-dependent spheroid morphologies.
• For the first time, a compact 3D spheroid was generated from the previously non-spheroid-forming SW48 cell line by optimizing matrix conditions and plate treatment.
• Treating standard multiwell plates with an anti-adherent solution offers a low-cost, scalable alternative to commercial cell-repellent plates for spheroid generation.
• Co-culture with colonic fibroblasts reproduced tumor-stroma interactions within spheroids, enhancing their biological relevance for drug testing.
• The authors recommend adopting standardized 3D culture protocols to strengthen reproducibility and comparability in preclinical colorectal cancer research.