iPSC-Derived Human Hepatocytes (WT) - Definigen

Hepatocyte WT

DefiniGEN hepatocytes are functionally mature hepatocytes which can be plated and functionally maintained over a +20 day window with similar function to primary human hepatocytes PHH.
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iPSC-derived hepatocytes form tight junctions and typical hepatocyte cobblestone morphology
Product Overview

The cells display similar functional characteristics of primary human hepatocytes, and their functionality remains stable over a prolonged period of time in culture making them ideal for drug discovery, drug metabolism, and toxicology-related studies. These hiPSC-derived hepatocytes express key hepatitis markers such as CD81, SR-B1, Claudin-1 and Occludin at similar levels to PHH making them an effective model for hepatitis lifecycle studies.

  • Highly standardized cell product containing >98% human hepatocyte cells with consistent performance and biologically relevant data
  • Wild-type donor genetics and karyotype verified
  • Cells are functional and stable over a +20 day window
  • Cells display CYP450 induced activities
  • Cells express key hepatocyte proteins, including A1AT, ALB, HNF4a,
  • Cells secrete physiologically relevant levels of albumin and urea
  • Cells express a range of key hepatitis B & C markers
  • Cells demonstrate LDL uptake
Product Specification
Ordering information
Product ID Def-HEP WT
 Format Cryopreserved
Cell number 4-6 million cells
Genetic background Wild-type
Viability >70%
Pricing Request a quote

Technical Data
Hepatocyte cell morphology

When thawed and plated as a monolayer, Def-HEP cells form hepatocytes with characteristic cobblestone morphology and tight cell junctions

iPSC-derived Hepatocytes form tight junctions and typical hepatocyte cobblestone morphology
Figure 1. Overview of Def-HEP cell morphology. Def-HEP WT cells exhibit typical hepatocyte cobblestone morphology and bi-nucleation.

Hepatocyte maturation markers

QPCR analysis shows Def-HEP cells show key hepatocyte markers at similar levels to PHH. Functional characteristics including albumin secretion, A1AT production, glycogen storage and LDL uptake are also present.

Human hepatocytes exhibit functional characteristics including albumin secretion, glycogen storage and LDL uptake
Figure 2. Functional analysis of Def-HEP WT hepatocytes (A) Albumin secretion, 10x magnification (B) Glycogen storage disease shown by PAS staining (C) LDL cholesterol uptake shown by fluoresceinated LDL incorporation.
Human hepatocytes express key hepatic gene markers including A1AT and ALB.
Figure 3. Gene expression analysis demonstrates that Def-HEP express key hepatocyte markers at similar levels to PHH. AFP levels are extremely low in Def-HEP indicating that the cells have attained a functional mature status.
Extended culture time
Functional Window Of Use DefiniGEN Hepatocytes
Figure 4. Extended functional window of use with DefiniGEN hepatocytes. Cryopreserved Def-HEP cells are thawed, plated, and recovered over 7 days to ensure a functional hepatocyte monolayer forms. Subsequently the cells have a +20 day functional window to enable hepatitis disease modelling, and toxicology studies to be undertaken over a longer timeframe than is possible with PHH.
Multiple Inducible CYP450 Activities

Def-HEP cells display CYP450 induced activity profiles that are similar to PHH (CYP1A2 EROD assay, inducer - omeprazole), (CYP3A4 PGlo assay, inducer - rifampicin).

CYP450 activity profiles in DefiniGEN hepatocytes
Figure 5. Multiple CYP activities of cryopreserved Def-HEP hepatocyte cells. The results show Def-HEP cells have comparable CYP activity to PHH and induced activity profiles that are highly similar to PHH (CYP1a2 EROD assay, inducer – omeprazole), (CYP3A4 PGlo assay, inducer – rifampicin).
Hepatitis marker analysis

Hepatitis marker analysis
Figure 6. Gene expression analysis of key Hepatitis markers. The analysis indicates the presence of key hepatitis markers in Def-HEP cells including NTCP, Occludin, SR-B1, CD81 and CLDN7. (HepG2 cells do not express all key hepatitis markers).

Mitochondrial Toxicity

As assessed by dosing Def-HEP WT with valinomycin (0.3-600nM) and papverine (0.03 – 60µM) for 48 hrs both compounds caused significant mitochondrial toxicity with concentration ranges recognized in literature. Cell viability was assessed via the CellTiter 96® Aqueous Non-Radioactive Cell. Both valinomycin and papaverine caused significant mitochondrial toxicity

Mitochondrial toxicity analysis DefiniGEN hepatocytes
Figure 7. Typical Def-HEP WT mitochondrial toxicology drug response to valinomycin and papverine.