Produção de esferoides de células tumorais de câncer de bexiga num biorreator de vórtice de Taylor

Abstract

Two-dimensional (2D) cell culture has been the common platform for evaluating anticancer drugs. However, three-dimensional (3D) cell cultures, such as multicellular tumor spheroids (MCTS), best resemble tumor in vivo properties, being potentially more suitable for preclinical evaluation of drug candidates. However, this shift from 2D to 3D on the high-throughput screening platform will only be possible if mass production of this culture becomes possible. This study aims to evaluate a new Taylor vortex bioreactor (TVB) for the production of MCTS using the RT4 bladder cancer cell line. For a robust analysis, a preliminary study was carried out by full factorial design of the independent variables agitation and inoculum density, resulting in 4 combinations and a central point. With the results obtained it was possible to make a more directed central composite design of the two variables, resulting in 9 different combinations and a triplicate in the central point. The response variables analyzed related to the spheroids produced were: mean diameter, standard deviation of this diameter, quantity (number), productivity, and circularity. Cultures were performed on a TVB with a working volume of 100 mL using McCoy 5A medium in a 37 °C incubator and 5-10% CO2. The results allowed the identification of values of the two independent variables that lead to cell growth with low lethality, high spheroid productivity and a wide range of diameters that could meet several technological demands in the evaluation of drug efficacy and toxicity. The circularity and variability of spheroid diameter, although considered good, need to be improved for new technological applications in medicine and pharmacy. In summary, it can be said that the use of TVB for the production of large-scale MCTS is a very promising platform.