Evaluation of different models of tumor spheroid generation with CAL27 cells

Abstract

In cancer research, three-dimensional cell culture models have become useful tools for investigating the tumor microenvironment, cancer stages, and the search for new therapeutic targets. In this sense, the aim of this study was to contribute with a tumor spheroid model of squamous cell carcinoma (CAL27) to be used for studies of this type of cancer and new therapeutic alternatives. For this, two methods of spheroid generation were tested, a 96-well plate model coated with agarose and a cylindrical microwell model developed using 9x9 size micromolds with 800 μm diameter that fit into 24-well plates (3D Petri Dish®). The microwell generation model showed more promising results, generating spheroids up to 183.97 ± 15 µm in diameter. Although the analysis of spheroid size and growth kinetics was superior to other assays, the marking of the necrotic center produced essential images for confirmation of the generation of the three-dimensional model, as well as the results of the cytotoxicity assay. The results presented here contribute to the progress of the use of 3D cell culture systems for the study and search for responses to agents that may act on squamous cell carcinoma and provide a viable alternative for cancer research laboratories.