HepG2-CYP™ Cell Panel

HepG2 is one of the most commonly used cell systems for in vitro screening, but HepG2 cells have low expression and activity of CYP enzymes which leads to missed or underestimated cytotoxicity of test compounds and/or metabolites

We have assembled a panel of HepG2 transgenic cell lines that each stably express one of the major human CYPs (originally published by Takeda Pharmaceutical Company Limited, Osaka, Japan) and utilized gene editing technology to create a CYP reductase (POR) knockout HepG2 cell line.

Used in appropriate assay panels, these cells can address a variety of questions pertaining to CYP mediated metabolism and drug/drug interaction

• CYP metabolism phenotyping can be accomplished directly with minimal or no use of inhibitors
• Direct or indirect determination of CYP inhibition
• CYP specific cytotoxicity

CYP activity in HepG2 WT, HepG2-CYP transgenic and HepG2-CYP reductase KO (POR) cells. CYP activity was measured  with P450 –Glo™ assay kits. Specific CYP activity in transgenic cells was confirmed with CYP-selective competitive inhibitors, namely 5 µM α-naphthoflavone (CYP1A2),  1 µM clopidogrel (CYP2B6),  2 µM sulfaphenazole (CYP2C9),  and 1 µM ketoconazole (CYP3A4).

The HepG2-CYP cell lines are available as a panel or can be purchased individually

Hera provides in vitro screening services using the HepG2-CYP panel as well other cell lines including: primary hepatocytes, HepaRG, Upcyte hepatocytes, Corning HepatoCells

• Cytotoxicity – cell viability (MTT & ATP)
• Metabolic stability & metabolite profiling
• CYP phenotyping and drug-drug interaction studies

Custom gene editing services (knockout, knock-in and transgenic) to improve metabolic capability
of any cell line as well as further modifications to any of the cell lines from the HepG2-CYP panel

• Establishment of the transformants expressing human cytochrome P450 subtypes in HepG2, and their applications on drug metabolism and toxicology (Hashizume et al, 2010)
• Advantages of Human Hepatocyte-Derived Transformants Expressing a Series of Human Cytochrome P450 Isoforms for Genotoxicity Examination (Yoshitomi et al, 2001)
• In vitro micronucleus test in HepG2 transformants expressing a series of human cytochrome P450 isoforms with chemicals requiring metabolic activation (Hashizume et al, 2009)