In preclinical validation processes in drug research, monolayer cell cultures are still mainly used. However, 2D cultures can only mimic the conditions of physiological tissue to a limited extent, while in vivo cells are able to three-dimensional to interact. Therefore, results from 2D cultures can often have limited relevance to clinical effectiveness and contribute to a high rate of attrition in the drug discovery process. The use of spheroid cultures is considered a better method for more predictable in vitro screening assays for preclinical drug research, especially in cancer research.
In spheroid cultures, cells grow in a three-dimensional system with zones of cellular heterogeneity and nutrient and oxygen gradients to better represent the in vivo tumor microenvironment. Comparisons of spheroid cultures with monolayer 2D cultures demonstrated functional differences in tumor cell lines, such as changes in protein expression, phosphorylation patterns, and responsiveness to inhibitory molecules.
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