Table 1.

Differences in two-dimensional vs three-dimensional cell culture models

Type of culture	2D	3D	Ref.
In vivo-like	Do not mimic the natural structure of the tissue or tumor mass	In vivo tissues and organs are in 3D form	Takai et al[102]
Proliferation	Tumor cells were grown in monolayer faster than in 3D spheroids	Similar to the situation in vivo	Lv et al[11]
Polarity	Partial polarization	More accurate depiction of cell polarization	Antoni et al[18]
Cell morphology	Sheet-like, flat, and stretched cells in monolayer	Form aggregate/spheroid structures	Breslin et al[103]
Stiffness	High stiffness (approximately 3 × 109 Pa)	Low stiffness (> 4000 Pa)	Krausz et al[104]
Cell-cell interaction	Limited cell-cell and cell-extracellular matrix interactions and no “niches”	In vivo-like, proper interactions of cell-cell and cell-extracellular matrix, environmental “niches” are created	Lv et al[11], Kang et al[105]
Gene/protein expression	Changes in gene expression, mRNA splicing, topology, and biochemistry of cells, often display differential gene/protein levels compared with in vivo models	Expression of genes and proteins in vivo is relevantly presented in 3D models	Bingel et al[92], Ravi et al[106]
Drug responses	Lack of correlation between 2D monolayer cell cultures and human tumors in drug testing.	Tumor cells in 3D culture showed drug resistance patterns similar to those observed in patients	Lv et al[11], Bingel et al[92]
The culture formation	From minutes to a few hours	From a few hours to a few days	Dai et al[33]
Quality of culture	High performance, reproducibility, long-term culture, easy to interpret, simplicity of culture	Worse performance and reproducibility, difficult to interpret, cultures are more difficult to carry out	Hickman et al[107]
Access to essential compounds	Unlimited access to oxygen, nutrients, metabolites, and signaling molecules (in contrast to in vivo)	Variable access to oxygen, nutrients, metabolites, and signaling molecules (similar to in vivo)	Pampaloni et al[108], Senkowski et al[30]
Cost during maintenance of a culture	Cheap, commercially available tests and media	More expensive, more time-consuming, fewer commercially available tests	Friedrich et al[35]