Comparison of two and three-dimensional in vitro tumor models
Júlia Crispim da Fontoura¹,²*, Pítia F. Ledur ¹,², Christian Viezzer¹,², Cristina B. C. Bonorino¹,².
¹Programa de Pós-Graduação em Biologia Celular e Molecular, Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS), Brazil, ²Instituto de Pesquisas Biomédicas, Hospital São Lucas da PUCRS, Brazil
There is a great gap between in vitro and in vivo models when considering screening of new drug candidates, generating an excessive use of animals. Poor in vitro models may lead to discarding potential therapeutic molecules due to inadequate transposition of the complexity seen in animal models. In order to solve this problem, three-dimensional (3D) in vitro models were developed, which demonstrates more accurately how the tumor microenvironment in vivo works. The aim of this study is to compare a developed 3D tumor model with an established 3D and a 2D one, considering the interactions that occur in the tumor microenvironment. Firstly, we standardized the use of a biodegradable electrospun membrane, using B16F10 melanoma cells. After 14 days of culture, membranes were analyzed through Scanning Electron Microscopy (SEM), where it was possible to distinguish 3 different morphologies: epithelial (cuboid), mesenchymal (star-like) and spheroid (group of cells). Then co-cultures were done with fibroblasts (NIH3T3), B16F10 and macrophages (RAW), where the formation of spheroids, as well as production of microvesicles was observed on 3D cultures, but not on 2D. Finally, experiments to see proliferation after drug treatment are under way, where different tumor cell lines will be cultured on this membrane, on Geltrex® and on a 2D model. In order to compare these cultures, RNA expression and cytokine production will also be evaluated. Data obtained in this system will increase knowledge on cell interaction and macrophage influence on tumor cells. Moreover, a future comparison of antitumor drug activity in these models will generate data to support the validation of such method, and may improve the initial stage of drug screening and decrease the use of experimental animals in the future.
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