Cytochrome P450 1A1/2, 2B6 and 3A4 HepaRG Cell-Based Biosensors to Monitor Hepatocyte Differentiation, Drug Metabolism and Toxicity
Abstract
:1. Introduction
2. Materials and Methods
2.1. Reagents
2.2. HepaRG Cell Culture
2.3. GFP Reporter Dene Constructs.
2.4. Lentiviral Infection and Selection of Biosensor HepaRG Cells.
2.5. Immunoblotting Blotting
2.6. RNA Expression by RetroTranscription Quantitative Polymerase Chain Reaction
2.7. Fluorescence Microscopy and Flow Cytometry Analyses.
2.8. Statistical Analyses
3. Results
3.1. Subsection
3.1.1. Expression of Phase I and II Enzymes in HepaRG Hepatocyte-Like Cells.
3.1.2. Establishment of Biosensor HepaRG Cells
3.1.3. Induction of GFP Expression in Biosensor HepaRG Cells by Xenobiotics
3.1.4. Detection of Selective Hepatocyte-Like Cytotoxicity
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Forward | Reverse | |
---|---|---|
CYP1A1 | TCTTCCTTCGTCCCCTTCAC | ACACCTTGTCGATAGCACCA |
CYP2B6 | TTCCTACTGCTTCCGTCTATC | GTGCAGAATCCCACAGCTCA |
CYP2E1 | TTGAAGCCTCTCGTTGACCC | CGTGGTGGGATACAGCCA |
CYP3A4 | CTTCATCCAATGGACTGCATAAA | TCCCAAGTATAACACTCTACACACACA |
CK19 | TTTGAGACGGAACAGGCTCT | AATCCACCTCCACACTGACC |
GFP | ACAACAGCCACAACGTCTAT | GGGTGTTCTGCTGGTAGTG |
TBP | GAGCTGTGATGTGAAGTTTCC | TCTGGGTTTGATCATTCTGTA |
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Vlach, M.; Quesnot, N.; Dubois-Pot-Schneider, H.; Ribault, C.; Verres, Y.; Petitjean, K.; Rauch, C.; Morel, F.; Robin, M.-A.; Corlu, A.; et al. Cytochrome P450 1A1/2, 2B6 and 3A4 HepaRG Cell-Based Biosensors to Monitor Hepatocyte Differentiation, Drug Metabolism and Toxicity. Sensors 2019, 19, 2245. https://doi.org/10.3390/s19102245
Vlach M, Quesnot N, Dubois-Pot-Schneider H, Ribault C, Verres Y, Petitjean K, Rauch C, Morel F, Robin M-A, Corlu A, et al. Cytochrome P450 1A1/2, 2B6 and 3A4 HepaRG Cell-Based Biosensors to Monitor Hepatocyte Differentiation, Drug Metabolism and Toxicity. Sensors. 2019; 19(10):2245. https://doi.org/10.3390/s19102245
Chicago/Turabian StyleVlach, Manuel, Nicolas Quesnot, Hélène Dubois-Pot-Schneider, Catherine Ribault, Yann Verres, Kilian Petitjean, Claudine Rauch, Fabrice Morel, Marie-Anne Robin, Anne Corlu, and et al. 2019. "Cytochrome P450 1A1/2, 2B6 and 3A4 HepaRG Cell-Based Biosensors to Monitor Hepatocyte Differentiation, Drug Metabolism and Toxicity" Sensors 19, no. 10: 2245. https://doi.org/10.3390/s19102245
APA StyleVlach, M., Quesnot, N., Dubois-Pot-Schneider, H., Ribault, C., Verres, Y., Petitjean, K., Rauch, C., Morel, F., Robin, M.-A., Corlu, A., & Loyer, P. (2019). Cytochrome P450 1A1/2, 2B6 and 3A4 HepaRG Cell-Based Biosensors to Monitor Hepatocyte Differentiation, Drug Metabolism and Toxicity. Sensors, 19(10), 2245. https://doi.org/10.3390/s19102245