Unlocking the Potential of Spheroids in Personalized Medicine: Systematic Review of Seeding Methodologies

Three-dimensional (3D) spheroid models have revolutionized in vitro cancer research by offering more physiologically relevant alternatives to traditional two-dimensional (2D) cultures. This systematic review evaluates methods for spheroid formation and cellular sources used, highlighting diverse applications and preferences in the field. The five most investigated cancer origins for spheroid seeding are breast, colon, lung, ovary, and brain cancers, reflecting their clinical importance and research focus. Among seeding methodologies, forced-floating and scaffold-based methods predominate, demonstrating reliability and versatility in spheroid generation. Other techniques, including microfluidics, bioprinting, hanging drop and suspension culture also play significant roles, each with distinct advantages and limitations. This review underscores the growing adoption of spheroid models and the need for standardization in methodologies to enhance reproducibility and translational potential in cancer research.