Novel in vitro liver metabolism models and their applications progress

doi:10.12092/j.issn.1009-2501.2026.07.014

Abstract

Abstract:

Studies of hepatic drug metabolism are crucial for drug development, clinical dosing guidance, and assessment of drug safety. Although traditional in vitro liver models have been widely used, their limitations in maintaining hepatic function, predictive accuracy, and physiological fidelity have spurred the development of various advanced in vitro liver models. Previous reviews have focused on the technologies used for constructing novel models or on comparative evaluations of their performance, whereas systematic analyses of the application characteristics of different advanced models in drug metabolism research remain limited. Based on a comprehensive review of recent advanced in vitro liver models and the novel technologies they employ, this article further aligns model functional characteristics with specific drug-metabolism research scenarios and summarizes their performance in metabolic stability assessment, metabolite identification, and the evaluation of drug–drug interactions. This review aims to provide guidance for the rational selection and standardized application of in vitro models, thereby supporting preclinical research and clinical use of drugs.

Key words: liver, biotransformation, in vitro techniques, metabolic clearance rate, drug interaction

CLC Number:

R969.1

Sihan LU, Guanglin BAO, Jianguo SUN. Novel in vitro liver metabolism models and their applications progress[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2026, 31(7): 975-989.

Figures/Tables 3

References 110

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Model	Common cell sources	Cell culture techniques	Advantages	Limitations
Sandwich culture model	PHH	Sandwich culture	Well-defined bile canaliculi, sustained metabolic function for extended periods, suitable for hepatobiliary drug-excretion and transporter studies	Limited physiological relevance in two-dimensional formats, limited maintenance of metabolic function
Spheroid model	PHH, iPSC, HepG2, HepaRG	ULA plate culture, hanging drop culture, hydrogel encapsulation, micropatterned plate culture, cell co-culture	High-throughput; long-term maintenance of metabolic function, physiological polarity and extracellular matrix, suitable for metabolism studies of low-clearance drugs	Prone to necrotic-core formation, heterogeneous spheroid formation, challenging real-time monitoring of spheroid interiors
Liver organoid model	Liver tissue, iPSC	Cell co-culture, hydrogel encapsulation, scaffold engineering	High fidelity to organ biology, long-term maintenance of metabolic function, suitable for studies of inter-individual variability	Prone to necrotic-core formation, complex culture procedures, high cost
Liver-on-a-chip	PHH, iPSC, HepG2, HepaRG	Microfluidic integration, micropatterned culture, cell co-culture, three-dimensional scaffold culture	High physiological relevance, Reduced susceptibility to central necrosis, long-term maintenance of metabolic function, real-time online monitoring, precise control of microenvironmental physiological parameters, suitable for complex metabolic studies	High technical barrier, high cost

Model	Common cell sources	Cell culture techniques	Advantages	Limitations
Sandwich culture model	PHH	Sandwich culture	Well-defined bile canaliculi, sustained metabolic function for extended periods, suitable for hepatobiliary drug-excretion and transporter studies	Limited physiological relevance in two-dimensional formats, limited maintenance of metabolic function
Spheroid model	PHH, iPSC, HepG2, HepaRG	ULA plate culture, hanging drop culture, hydrogel encapsulation, micropatterned plate culture, cell co-culture	High-throughput; long-term maintenance of metabolic function, physiological polarity and extracellular matrix, suitable for metabolism studies of low-clearance drugs	Prone to necrotic-core formation, heterogeneous spheroid formation, challenging real-time monitoring of spheroid interiors
Liver organoid model	Liver tissue, iPSC	Cell co-culture, hydrogel encapsulation, scaffold engineering	High fidelity to organ biology, long-term maintenance of metabolic function, suitable for studies of inter-individual variability	Prone to necrotic-core formation, complex culture procedures, high cost
Liver-on-a-chip	PHH, iPSC, HepG2, HepaRG	Microfluidic integration, micropatterned culture, cell co-culture, three-dimensional scaffold culture	High physiological relevance, Reduced susceptibility to central necrosis, long-term maintenance of metabolic function, real-time online monitoring, precise control of microenvironmental physiological parameters, suitable for complex metabolic studies	High technical barrier, high cost

Compound	Fold error
Compound	HepaRG	SCC	Suspension	MPCC	Sph
Diazepam	0.2	0.6	NA	1.1	0.8
Etodolac	0.8	0.7	0.3	0.5	1.6
Glipizide	0.2	0.3	0.8	0.4	0.5
Ketoprofen	0.9	1.6	0.7	2.2	0.9
Ondansetron	1.4	1.0	1.8	2.5	1.8
Theophylline	NA	ND	NA	1.7	3.5
Tolbutamide	ND	1.9	4.9	1.2	2.3
Warfarin	1.3	1.0	NA	2.3	1.4

Compound	Fold error
Compound	HepaRG	SCC	Suspension	MPCC	Sph
Diazepam	0.2	0.6	NA	1.1	0.8
Etodolac	0.8	0.7	0.3	0.5	1.6
Glipizide	0.2	0.3	0.8	0.4	0.5
Ketoprofen	0.9	1.6	0.7	2.2	0.9
Ondansetron	1.4	1.0	1.8	2.5	1.8
Theophylline	NA	ND	NA	1.7	3.5
Tolbutamide	ND	1.9	4.9	1.2	2.3
Warfarin	1.3	1.0	NA	2.3	1.4

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