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 ID:
- Def-HEP WT
- 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.
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.
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.
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).
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).
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.
Figure 6. Confirmation of absence of the Crabtree effect in Def-HEP WT.
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 7. Typical Def-HEP WT mitochondrial toxicology drug response to valinomycin and papverine.
Brochure: Definigen Hepatocytes Wild Type
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