Currently, there is a large demand on drug production for oncology, mainly for bladder cancer treatment due to its high incidence. The development of a new drug evolves extensive analysis of effectiveness and safety during pre-clinical phase. Traditionally, cells cultured in monolayer, or 2D, have been applied for in vitro assays. However this model does not represent the best way to evaluate new compounds once it does not mimic the tumor microenvironment. The culture of tumor cells in tri-dimensional (3D) conditions is a method that has been widely investigated. Multicellular spheroids are the most used 3D models because the diffusion profile of oxygen, nutrients, wastes and drugs are more reliable comparing to in vivo models. The presence of a hypoxia core within the spheroid is associated to resistance mechanisms to some treatments, as observed in solid tumors. 3D spheroids are produced by different techniques and it's still unknown of which one is more efficient in terms of robustness, reliability, and versatility. One of them applies dynamic systems that yield a high number of spheroids with relatively uniform diameters that could be scaled up. This study will focus in the evaluation of the 3D culture of the cell line RT4 in stirred systems to establishment a 3D bladder cancer model to be applied in cytotoxicity assays and drug screening.
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