Optimized human cell products for research
and drug discovery

Human Hepatocytes Wild Type

DefiniGEN Human Hepatocytes Wild Type

DefiniGEN human hepatocyte cells are high functionality cryopreserved cell products displaying many of the functional characteristics of primary human cells. When grown in monolayer the cells display typical hepatocyte tight cell junction cobblestone morphology and  bi-nucleated cells are also present. Key hepatocyte functions including albumin production, glycogen storage, CYP450 mRNA expression and catalytic activities are observed at levels comparable to primary human hepatocytes. In terms of general metabolism DefiniGEN hepatocytes generate ATP via mitochondrial oxidative phosphorylation and thus are suitable for mitochondrial toxicology applications and they do not exhibit the Crabtree Effect, the disadvantageous phenomenon observed in immortalized liver cell lines which generate their ATP via glycolysis. Following thaw and recovery DefiniGEN hepatocytes have a 15-20 day effective window of use making them effective models for hepatitis lifecycle studies and the cells express key hepatitis markers such as CD81, SR-B1, Claudin-1 and occulin at similar levels to primary human hepatocytes.

Product Specification

Product ID:
Cell number:
Typically 6 million cells per unit
None (Healthy Donor)

Hepatocyte Cell Morphology

The Def-HEP cell products are highly functional human hepatocytes. When thawed and plated as a monolayer, Def-HEP cells form hepatocytes with cobblestone morphology and tight cell junctions.

Hepatocyte WT Morphology

Figure 1. Overview of Def-HEP cell morphology. Def-HEP WT cells exhibit typical hepatocyte cobblestone morphology and bi-nucleation.

Hepatocyte Maturation Markers

Def-HEP cells display the functional characteristics of primary human hepatocytes (PHH) including albumin secretion, A1AT production, glycogen storage and LDL uptake (Figure 2). Accordingly they should be handled in a similar fashion to Primary Human Hepatocytes.

Hepatocyte Maturation Markers

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. 

Hepatocyte Maturation Marker Analysis

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

Hepatocyte Marker Analysis

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.

Multiple Inducible CYP450 Activities

Def-HEP cells display CYP450 induced activity profiles that are similar to PHH (CYP1a2 EROD assay, inducer – omeprazole), (CYP450 PGlo assay, inducer – rifampicin).

Multiple Inducible CYP450 Activity

Multiple Inducible CYP450 Activity


Figure 4. Multiple CYP activites 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), (CYP450 PGlo assay, inducer – rifampicin).

Hepatitis Marker Analysis

Gene expression analysis demonstrates that multiple batches of Def-HEP cells reproducibly express key hepatitis markers including CD81, SR-B1, Claudin-1 and occulin. The levels of markers observed are extremely similar to PHH (HepG2 cells do not express all key hepatitis markers).

Gene Expression Analysis

Figure 5. Gene expression analysis demonstrating the presence of key Hepatitis C markers in Def-HEP including SR-B1, CD81, Claudin-1 and Occludin.

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.

Absence Of Crabtree Effect

Figure 6. 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

Mitochondrial Toxicity

Figure 7. Typical Def-HEP WT mitochondrial toxicology drug response to valinomycin and papverine.


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