Hepatocyte WT

Ready to order?
UK +44 (0)1223 497106
USA 617-674-3260

DefiniGEN provide mature hepatocytes derived from hIPSC. 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

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 shown by PAS staining (C) LDL cholesterol uptake shown by fluoresceinated LDL incorporation.



Human hepatocytes express key hepatic gene markers including AAT 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 the cells have attained a functional mature status.

Extended culture time


Figure 4. 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).



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


Figure 6. Gene expression analysis demonstrates that multiple batches of Def-HEP cells reproducibly express key hepatitis markers including CD81, SR-B1, Claudin-1 and Occludin. The levels of markers observed are similar to PHH (Hep G2 cells do not express all key hepatitis markers)

Metabolism – Absence Of Crabtree Effect

Dosing Def-HEP WT with +/- glucose media does not alter the hepatotoxic effect of chlorpromazine as assessed via the MTT cell proliferation assay. Therefore, it is highly unlikely that our cells exhibit the Crabtree effect, in contrast to immortalized lines.


Figure 7. Confirmation of absence of the Crabtree effect in Def-HEP WT.

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


Figure 8. Typical Def-HEP WT mitochondrial toxicology drug response to valinomycin and papverine.

We recommend that you use our online system to identify the products of interest and their respective product codes. To order we will require the following information:

Product code(s) of the item(s) you wish to order (e.g. Def-HEP WT)
Your name, email address, title and organization
Shipping address and billing address with contact names
Purchase Order number if you have one
Telephone number

For further information please contact us: