Dear Karim,

The attached paper covers the answer to your question. I have copied the material and methods section for quick view.In this section you can find the company where the required cell line was purchased.

Materials and Methods

Cell Culture.

The human liver cell line THLE2, which was derived from primary normal liver epithelial cells, was purchased from the American Type Culture Collection (Manassas, VA). THLE2 cells were cultured in LHC-8 medium (Invitrogen, Carlsbad, CA) supplemented with 70 ng/ml phosphoethanolamine, 5 ng/ml epidermal growth factor, 10% fetal bovine serum (Atlanta Biologicals, Lawrenceville, GA), and the antibiotics penicillin (50 U/ml), and streptomycin (50 μg/ml) (Sigma-Aldrich, St. Louis, MO). Human hepatoma cell lines HepG2, Hep3B, Huh7, and SK-Hep-1 (American Type Culture Collection) were cultured in Dulbecco's modified Eagle's medium (Sigma-Aldrich) supplemented with 10% fetal bovine serum, 2 mM l-glutamine, 50 U/ml penicillin, and 50 μg/ml streptomycin. The passage number was less than 10 for all experiments performed in each cell type. Cells were seeded onto 60 × 15-mm cell culture dishes at a cell density of 5 × 105 in 5 ml of culture media and were maintained at 37°C in a humidified atmosphere with 5% CO2 until they were 70 to 80% confluent (cell confluence was evaluated by visual observation using an optical microscope).

Human primary hepatocytes from anonymous donors were obtained through the Liver Tissue Cell Distribution System (Pittsburgh, PA). Donor information is listed in Supplemental Table 1. Hepatocytes were isolated by a three-step collagenase perfusion as described previously (Strom et al., 1996). Upon arrival, the shipping medium was removed and replaced with serum-free hepatocyte maintenance medium supplemented with insulin and GA-1000 using HMM SingleQuots (Lonza Walkersville, Inc., Walkersville, MD). Primary hepatocytes were plated on collagen in T-25 flasks containing approximately 106 cells. The cultured hepatocytes were incubated at 37°C in a humidified atmosphere of 5% CO2 for at least 12 h before harvesting. This project was approved by the Research Involving Human Subjects Committee of the U.S. Food and Drug Administration.

RNA Isolation.

Total RNA from hepatocytes or cell lines was isolated using an RNeasy system (QIAGEN, Valencia, CA). The yield of the extracted RNA was determined spectrophotometrically by measuring the optical density at 260 nm. The purity and quality of RNA were evaluated using an RNA 6000 LabChip on an 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA). High-quality RNA with RNA integrity numbers greater than 9.0 were used for the study.

Human Drug Metabolism RT2 Profiler PCR Array.

First Strand cDNA Synthesis Kits and human drug metabolism RT2 Profiler PCR arrays were obtained from SABiosciences. The human drug metabolism RT2Profiler PCR array contains a total of 251 drug metabolism genes and 5 endogenous control genes.

Real-Time Reverse Transcriptase-PCR.

For first-strand cDNA synthesis, 1 μg of total RNA was reverse-transcribed in a final volume of 20 μl of with random primers at 37°C for 60 min according to the manufacturer's instructions (SABiosciences). In brief, reverse transcriptase was inactivated by heating at 95°C for 5 min. The cDNA was diluted to 100 μl by adding RNase free water and stored at −20°C. The PCR was performed using an ABI 7900 instrument (Applied Biosystems, Foster City, CA). For one 96-well plate of the PCR array, 2450 μl of PCR Master Mix containing 1× PCR Master Mix and 98 μl of diluted cDNA was prepared, and an aliquot of 25 μl was added to each well. Three technical replicates were run for each RNA sample.

Data Normalization and Analysis.

Endogenous control genes, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and β-actin (ACTB) present on the PCR array were used for normalization. Each cycle threshold (Ct) was normalized to the average Ct of the two endogenous controls. The comparative ΔCt method was used to calculate relative quantification of gene expression.

Sensitivity Detection and Identification of Differentially Expressed Genes.

PCR array quantification was based on the Ct number. A gene was considered not detectable when Ct >32. A list of differentially expressed genes was identified using a two-tailed t test. The criteria were p < 0.05 and a mean difference ≥2-fold. The statistical calculations were based on ΔCt values.

Hoping this will be helpful,

Rafik

You could try HepG2 cell line from ATCC. Here is the link ....

http://www.atcc.org/products/all/HB-8065.aspx

Hope this helps!!!

HepaRG cells, "Upcyted" hepatocytes. HepG2 in our hands shows low/no expression of P450 enzymes.

I am not exactly sure what you mean by your question "expressing a P450 enzyme". Most cells express more than one, in fact multiple P450s, at various levels so if you are looking specifically at the activity of one enzyme then bactosomes will be a better option or a combination of cell lines with different expression patterns. To the best of my knowledge, and own work, there are no stable human cell line transfectants for Cytochrome P450. The only one we know that creates stable expression is the hamster cell line CHO. There is one publication from a Chinese group about a stable MCF7 cell line transfected with CYP1A1, however when we contacted them they said stable for a month..... so NOT stable.

The following paper may help.

Long-term culture and expansion of Primary human hepatocytes

Levy etal., Nat. Biotech December 2015.

I think ABM (www.abmgood.com) have HEK293 cells expressing variants of P450 available soon.