The creation of cell cultures for scientific purposes has made it possible to obtain new knowledge and consequently to make discoveries in the field of cell biology or biophysics. In vitro studies allow for the observation of cell lines as well as interactions with introduced substances or materials. They have an invaluable contribution to the development of nanomedicine, which is attracted a lot of interest. Gold nanoparticles (GNPs) are particularly popular and promising, especially in terms of cancer therapy. This is due to the specific (e.g. electrical, magnetic, optical, mechanical) properties of nanoparticles, significantly different from gold particles of macro size. Unfortunately, the results of in vitro studies are sometimes inconsistent with in vivo studies. Nanoparticles that work well at the cellular level are not always as effective in animal models. The reason for this is the multiplicity of complex metabolic processes occurring in the body during in vivo studies. Most cell studies are performed on two-dimensional structures that approximate real-world conditions. Currently, none of the in vitro techniques is able to reflect the identical physiological conditions prevailing in animal models. However, modern science can map them more precisely using 3D culture, which is much more complex and less efficient. When designing new studies, the advantages and disadvantages of each of the mentioned cell culture methods should be considered. The purpose of this publication is to present the differences between two-dimensional and three-dimensional (3D) cell cultures, taking into account the use of gold nanoparticles.
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