Table 1.

Advantages and drawbacks of cultivation systems for hepatocytes.

Cultivation	Advantages	Disadvantages	Reference
2D
Monolayer	Gold standard for drug metabolism and toxicity	Rapid loss of morphology and cell polarity	[8,9,15]
	Ideal for testing interindividual and interspecies differences in metabolism	Rapid loss of drug metabolizing capability, decrease of albumin production and cell-cell interaction)
	Maintenance of key functions as carbohydrate metabolism and plasma protein synthesis (24–72 h)	Limited availability
Co-culture	Improved functionality of all cell types	No real standard established	[8,16,17,18,19]
	Increased expression of phase I and phase II enzymes	High variability between different laboratories
	Maintenance of cell morphology
	Inducibility of CYPs
3D
Hydrogels, scaffolds scaffold-free	Long-term maintenance of liver-specific functions	Lack of established standards	[8,9,15,20,21,22,23,24,25]
	Increased sensitivity towards drugs	Not adjusted to high throughput
	Long-term expression of phase I and phase II enzymes	Cell recovery for further analysis is difficult
Co-culture with liver-drived cell types	Improved expression of phase I and phase II enzymes, Including inducibility by drugs	No real standard established	[17,26,27]
		Not adjusted to high throughput, high variability regarding cell viability and differentiation
	Maintenance of cell polarity, cell-cell contacts and bile canaliculi
	Mimicks in vitro architecture
	Longer cell viability
Microfluidic devices	Sustained liver like cell functionality and increased liver specific functions	No standardized system available so far	[8,9,15]
		Not adjusted to high throughput
	Precisely adjusted flow/drug concentrations
	Enable microscopic examination
	Formation of a sinusoid-like shape (HepaChip®)
	Fast differentiation of the cells after flow induction