In vitro screening for anti-cancer agents currently uses mainly cell lines in 2D culture. It is generally
assumed that 3D culture, namely spheroids, represents physiologically more relevant models for tumors. Unfortunately,
drug testing in spheroids is not as easy and reproducible as in 2D culture because there are factors that
limit the universal use of spheroids as screening platforms. Technical problems in the generation of uniform spheroids,
cell/tumor-specific differences in the ability to form spheroids, and more complex readout parameters are
the main reasons for differences between spheroid data. The review discusses requirements for cancer spheroids
to be representative models, suitable methodologies to generate spheroids for the screening and readout parameters
for the evaluation of anti-cancer agents.
Keywords: Drug screening, cell culture, 3D culture, cancer spheroids, anti-cancer drugs, 2D culture.
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