Table 1.

Microphysiological Technologies Used to Model the Hepatic Microenvironment

Representative Image	Platform	Scaffold/Format	Co-culture or integrated with other organ tissues	Validated functions of phase I/II and transporters	Other liver-specific functions	Methods used to assess hepatotoxicity
	HepatoPac Hepregen (32-33)	Micropatterned 24- and 96-well plates	Primary hepatocytes islands co-cultured with 3T3 fibroblasts	Metabolite measurements: CYP1A: Ethoxy-resorufin; CYP2A6: Coumarin; CYP2B6: Bupropion; CYP3A4: testosterone UGTs and SULTS: 7- hydroxycoumarin mRNA expression : phase I CYPs, FMO, MAO, OX, EH; phase II- UGTs, SULTs, NATs, GSTs, methyltransferases Transporters-MDR1, MRP3, OCT1, NTCP, BSEP canalicular flow by CMFDA fluorescent probe	Albumin (ELISA) and urea (colorimetric endpoint assay) secretion	MTT assay, ATP, and glutathione levels using Cell Titer-Glo and GSH-Glo luminescent kits (Promega)
	RegeneMed Liver Transwell RegenMed (35)	24-well transwell plate	Primary human or rat hepatocytes with hepatic NPCs,including vascular and bile duct endothelial cells, Kupffer cells and hepatic stellate cells	Phase I activity: CYP1A1, 2C9, and 3A4 by P450-Glo assays (Promega) Basolateral transporter uptake measured by isotope labeled tracer - 3H-labeled estrone-3-sulphate	Albumin, urea, fibrinogen, transferrin production (ELISA); glycogen synthesis (isotope labeling) LPS induced IL-1β, IL- 6, IL-8, TNF-α, GM- CSF: multiplex-based ELISA	Levels of ATP and GSH, LDH releasing
	3D InSight™ Microtissues InSphero (36)	Spheroids in 96-well plate	Primary human hepatocytes with NPCs (Kupffer and endothelial cells)	CYPs activity/induction: CYP1A1, 2B6, 2C8,2C9,2C19,2D6 CYP3A4 and 2E1 (IHC staining) MDR1,BSEP (IHC staining)	Albumin secretion (ELISA) IL-6 and TNF-α releasing (ELISA)	ATP, and glutathione (GSH) levels using CellTiter-Glo and GSH-Glo luminescent kits (Promega) LDH releasing, mitochondrial activity
	3D print Liver Organovo	3D bioprinted liver tissues in a transwell plate	Primary human hepatocytes with NPCs	CYP450 enzymes 3A4 (mRNA expression levels and probe metabolite formation)	Albumin and transferrin production Immunologic: IL-6, GM-CSF,MCP-1 LPS stimulated TNF-α, IL-1β, IL-12p70, IL-10, IL-2, IL-13, and IL-4	LDH releasing, GSH and ATP level Gene/protein expression profiling and histological tissue assessment
	HμREL hepatic cultures Hurel (38)	mCCA biochip connected with pump mCCA biochip can integrate multiple organ tissues	Primary human hepatocytes with NPCs	mRNA expression and metabolite formation: CYP450 enzymes 1A2, 3A4, 2C19, SULT, UGT Metabolic clearance: Biliary efflux assay (LC-MS/MS) BSEP canaliculi (CMFDA)		MTT assay, ATP, and glutathione (GSH) levels using CellTiter- Glo and GSH-Glo luminescent kits (Promega), LIVE/DEAD stain (Thermo Fisher)
	LiverChip™ System CN Bio Innovations (39-41)	Multiwell plate platform with pneumatic pump	Primary hepatocytes co- cultures with NPCs and Kupffer cells	Metabolite formation: CYP450 enzymes 1A2, 2B6, 2C9, 2D6, 3A4		LIVE/DEAD stain (Thermo Fisher)
	CellAsic (42)		Primary human hepatocytes	Metabolite formation: CYP450 enzymes 1A1/2, 2C9, 3A4, UGT BSEP canaliculi (CMFDA)		LIVE/DEAD stain( fluorescent probe; Thermo Fisher)
	SQL-SAL University of Pittsburgh (43)	Nortis chip (Nortis, Inc.) with microfluidic flow	Primary human hepatocytes with cell lines derived from NPCs	Metabolite formation: CYP450 enzymes 1A, 2C9, 3A4 IHC staining: MDR1,MRP2, BSEP, BRCP	TNF-α releasing	LDH leakage, fluorescent protein ROS biosensors Stellate cell activation and migration
	IdMOC™ In Vitro ADMET Lab (72)	24 or 96-well plate format integrated with multiple organ	Primary human hepatocytes with 3T3 cells			MTT assay, ATP, and glutathione (GSH) levels Using CellTiter-Glo and GSH-Glo luminescent kits (Promega)
	Microcompartment hollow fiber BAL (37)	Hollow fiber	Primary human liver cells ( not pure hepatocytes)	Urea production , albumin synthesis , glucose secretion , and lactate production IHC: expressed cell marker of Kupffer cells)		LDH and AST releasing