Obtaining and characterization of spheroids using colon adenocarcinoma SW480 cells

Colon cancer has conventionally been studied using in vitro monolayer models to understand its biology and identify therapeutic agents; nevertheless, they do not represent tumor cells growing in three dimensions. This technical note presents a protocol to develop and characterize a three-dimensional model of spheroids of different sizes, formed with SW480 colon adenocarcinoma cells seeded on agarose
micromolds allowed to grow for up to 21 days. The spheroids were characterized by flow cytometry based on cell cycle, mitochondrial function, and apoptosis. Spheroids of 400 μm seeding size and 3 days of growth were found to represent a model possessing similar characteristics to those of tumors in vivo. An easy-toimplement method to evaluate therapeutic agents is proposed, previous to the use of animal models.