Table 3.

Advantages and disadvantages of commonly used 3D culture methodologies

	Advantages	Disadvantages
Spheroids/multicellular tumour spheroids (MCTS)	Easy to culture Mimic in vivo-like cell–cell and cell-ECM interactions Scalable and high-throughput (HTP) compliant Easily extracted for further experimentation	Size variability Limited diffusion if large Necrotic core formation Agglomeration Take time to form and show functionality
Scaffolds/hydrogels	Hydrogels: in vivo-like 3D interactions Used to study cell aggressiveness/metastatic potential Scaffolds: can be combined with functional tests	Hydrogels: size/shape variation. Hard to reproduce Difficult cell extraction Scaffolds: cells can flatten/adhere to scaffold Difficult materials can affect growth
Organoid/Tumouroid	In vivo-like architecture In vivo-like complexity Patient specific Replicate in vivo-like cell interactions	Complex to culture Variation Less amenable to HTS Needs much optimization/validation May lack certain cell types/vasculature
Liver-on-chip/microphysiological systems (MPS)	In vivo-like architecture In vivo-like chemical/physical gradients, microenvironment	Flow of medium may disrupt cells Difficult to adapt to HTS Lack vasculature
Explants/tumour explants	In vivo-like architecture In vivo-like complexity Useful in modelling disease	Variation between donors Difficult to obtain Complex to culture/expensive to maintain Lack long-term viability