Scientific background


The Caco-2 cell line is a continuous monolayer of heterogeneous human epithelial colorectal adenocarcinoma cells, developed by the Sloan-Kettering Institute for Cancer Research through research conducted by Dr. Jorgen Fogh. 

Although derived from a colon carcinoma, when cultured under specific conditions the cells become differentiated and polarized such that their phenotype, morphologically and functionally, resembles the enterocytes lining the small intestine. Caco-2 cells express tight junctions, microvilli, and a number of enzymes and transporters that are characteristic of such enterocytes: peptidases, esterases, P-glycoprotein, uptake transporters for amino acids, bile acids carboxylic acids, etc. 


The human intestinal Caco-2 cell line has been extensively used over the last twenty years as a model of the intestinal barrier.

Caco-2 cells are most commonly used not as individual cells, but as a confluent monolayer on a cell culture insert filter. When cultured in this format, the cells differentiate to form a polarized epithelial cell monolayer that provides a physical and biochemical barrier to the passage of ions and small molecules. The Caco-2 monolayer is widely used across the pharmaceutical industry as an in vitro model of the human small intestinal mucosa to predict the absorption of orally administered drugs. The correlation between the in vitro apparent permeability across Caco-2 monolayers and the in vivo fraction absorbed is well established.


Caco-2 monolayer properties allow it to be a reliable model for testing drugs in pre-clinical phases.

Before pharmaceutical companies start clinical trials on a drug, they conduct extensive pre-clinical studies. These involve in vitro (test tube or cell culture) and in vivo (animal) experiments using wide-ranging doses of the study drug to obtain preliminary efficacy, toxicity and pharmacokinetic information. CaCo-2 cell lines are a model of in vitro cell culture, and they are used as an alternative or a complement of animal testing. Such tests assist pharmaceutical companies to decide whether a drug candidate has scientific merit for further development as an investigational new drug.

New chemical entities (NCEs, also known as new molecular entities or NMEs) are compounds which emerge from the process of drug discovery. These will have promising activity against a particular biological target thought to be important in disease. However, little will be known about the safety, toxicity, pharmacokinetics and metabolism of this NCE in humans. It is the function of drug development to assess all of these parameters prior to human clinical trials.





DOT. Code of Federal Regulations, Title 49. 1.

FDA-Guidance. 2012.

Waiver of In Vivo Bioavailability and Bioequivalence Studies for Immediate-Release Solid Oral Dosage Forms Based on a Biopharmaceutics Classification System

Drug Interaction Studies — Study Design, Data Analysis, and Implications for Dosing and Labeling

The BCS, BDDCS, and Regulatory Guidances

EMA-Guidance. 2010. 

EMA-Guidance. 2012. 

IATA, I.A.T.A. 2014. Dangerous Goods Regulations 56th Edition.



Artursson P.
Epithelial transport of drugs in cell culture: I. A model for studying the passive diffusion of drugs over intestinal absorptive (Caco-2) cells
J. Pharm. Sci., 79 (1990), pp. 476–482

Artursson P. , Karlsson J.
Correlation between oral drug absorption in humans and apparent drug permeation coefficients in human intestinal epithelial (Caco-2) cells
Biochem. Biophys. Res. Commun., 175 (1991), pp. 880–885

Conradi R.A. , Hilgers A.R., Ho N.F.H., Burton P.S.
The influence of peptide structure on transport across Caco-2 cells
Pharm. Res., 8 (1991), pp. 1453–1460

Hidalgo I.J., Raub T.J., Borchardt R.T.
Characterization of the human colon carcinoma cell line (Caco-2) as a model system for intestinal epithelial permeability
Gastroenterology, 96 (1989), pp. 736–749

Hilgers A.R., Conradi R.A., Burton P.S.
Caco-2 cell monolayers as a model for drug transport across the intestinal mucosa
Pharm. Res., 7 (1990), pp. 902–910

Pinto M., Robine-Leon S., AppayM.D., Redinger M., Triadou N., Dussaulx E., Lacroix B., Simon-Assmann P., Haffen K., Fogh J., Zweibaum A.
Enterocyte-like differentiation and polarization of the human colon carcinoma cell line Caco-2 in culture
Biol. Cell.., 47 (1983), pp. 323–330



Peterson M.D., Mooseker M.S.
Characterization of the enterocyte-like brush border cytoskeleton of the C2BBe clones of the human intestinal cell line Caco-2

Woodcock S., Williamson I., Hassan I., Mackay M.
Isolation and characterization of clones from Caco-2 cell line displaying increased taurocholic acid transport
J. Cell Sci., 98 (1991), pp. 323–332

Caro I., Boulenc X., Rousset M., Meunier V., Bourrié M., Julian B., Joyeux H., Roques C., Berger Y., Zweibaum A., Fabre G.
Characterisation of a newly isolated Caco-2 clone (TC-7), as a model of transport processes and biotransformation of drugs
Int. Journal Phar., 116 (2) (1995), pp.147-158

Turco L., Catone T., Caloni F., Di Consiglio E., Testai E., Stammati A.
Caco-2/TC7 cell line characterization for intestinal absorption: how reliable is this in vitro model for the prediction of the oral dose fraction absorbed in human?



Abe, T., Unno, M., Onogawa, T., Tokui, T., Kondo, T.N., Nakagomi, R., Adachi, H., Fujiwara, K., Okabe, M., Suzuki, T., Nunoki, K., Sato, E., Kakyo, M., Nishio, T., Sugita, J., Asano, N., Tanemoto, M., Seki, M., Date, F., Ono, K., Kondo, Y., Shiiba, K., Suzuki, M., Ohtani, H., Shimosegawa, T., Iinuma, K., Nagura, H., Ito, S., and Matsuno, S. 2001. LST-2, a human liver-specific organic anion transporter, determines methotrexate sensitivity in gastrointestinal cancers. Gastroenterology 120:1689-1699.

Amy C. Ditewig, D.J.C., Michael J. Liguori, Yi Yang, and Eric A. G. Blomme. 2013. Transcriptomic evaluation of canine suspension-shipped and pre-plated hepatocytes: comparison to liver. Toxicology Mechanisms and Methods 23:479-490.

Anwer, M.S. and Stieger, B. 2014. Sodium-dependent bile salt transporters of the SLC10A transporter family: more than solute transporters. Pflugers Arch 466:77-89.

Arnaud Béduneau, C.T., Stéphane Fimbel, Yann Pellequer, Vincent Jannin, Frédéric Demarne, Alf Lamprecht. 2014. A tunable Caco-2/HT29-MTX co-culture model mimicking variable permeabilities of the human intestine obtained by an original seeding procedure. European Journal of Pharmaceutics and Biopharmaceutics 87:290-298.

Bailey, L.A., Hatton, D., Field, R., and Dickson, A.J. 2012. Determination of Chinese hamster ovary cell line stability and recombinant antibody expression during long-term culture. Biotechnology and bioengineering 109:2093-2103.

Beery, E., Rajnai, Z., Abonyi, T., Makai, I., Bansaghi, S., Erdo, F., Sziraki, I., Heredi-Szabo, K., Kis, E., Jani, M., Marki-Zay, J., Toth, G.K., and Krajcsi, P. ABCG2 modulates chlorothiazide permeability--in vitro-characterization of its interactions. Drug Metab Pharmacokinet 27:349-353.

Bentz, J., O'Connor, M.P., Bednarczyk, D., Coleman, J., Lee, C., Palm, J., Pak, Y.A., Perloff, E.S., Reyner, E., Balimane, P., Brannstrom, M., Chu, X., Funk, C., Guo, A., Hanna, I., Heredi-Szabo, K., Hillgren, K., Li, L., Hollnack-Pusch, E., Jamei, M., Lin, X., Mason, A.K., Neuhoff, S., Patel, A., Podila, L., Plise, E., Rajaraman, G., Salphati, L., Sands, E., Taub, M.E., Taur, J.S., Weitz, D., Wortelboer, H.M., Xia, C.Q., Xiao, G., Yabut, J., Yamagata, T., Zhang, L., and Ellens, H. Variability in P-glycoprotein inhibitory potency (IC(5)(0)) using various in vitro experimental systems: implications for universal digoxin drug-drug interaction risk assessment decision criteria. Drug Metab Dispos 41:1347-1366.

Blais, A., Aymard P Fau - Lacour, B., and Lacour, B. 1997. Paracellular calcium transport across Caco-2 and HT29 cell monolayers.

Briske-Anderson, M.J., Finley, J.W., and Newman, S.M. 1997. The influence of culture time and passage number on the morphological and physiological development of Caco-2 cells. Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine (New York, N.Y.) 214:248-257.

Burckhardt, G. and Burckhardt, B.C. 2011. In vitro and in vivo evidence of the importance of organic anion transporters (OATs) in drug therapy. Handb Exp Pharmacol 29-104.

Busch, A.E., Karbach, U., Miska, D., Gorboulev, V., Akhoundova, A., Volk, C., Arndt, P., Ulzheimer, J.C., Sonders, M.S., Baumann, C., Waldegger, S., Lang, F., and Koepsell, H. 1998. Human neurons express the polyspecific cation transporter hOCT2, which translocates monoamine neurotransmitters, amantadine, and memantine. Mol Pharmacol 54:342-352.

Choudhuri, S., Cherrington, N.J., Li, N., and Klaassen, C.D. 2003. Constitutive expression of various xenobiotic and endobiotic transporter mRNAs in the choroid plexus of rats. Drug Metab Dispos 31:1337-1345.

Coriell, L.L. 1979. Preservation, storage, and shipment. Methods in enzymology 58:29-36.

Cui, Y., Konig, J., and Keppler, D. 2001. Vectorial transport by double-transfected cells expressing the human uptake transporter SLC21A8 and the apical export pump ABCC2. Mol Pharmacol 60:934-943.

Dawson, P.A., Lan, T., and Rao, A. 2009. Bile acid transporters. J Lipid Res 50:2340-2357.

Dawson, S., Stahl, S., Paul, N., Barber, J., and Kenna, J.G. 2012. In vitro inhibition of the bile salt export pump correlates with risk of cholestatic drug-induced liver injury in humans. Drug Metab Dispos 40:130-138.

de Boussac, H., Orban, T.I., Varady, G., Tihanyi, B., Bacquet, C., Brozik, A., Varadi, A., Sarkadi, B., and Aranyi, T. Stimulus-induced expression of the ABCG2 multidrug transporter in HepG2 hepatocarcinoma model cells involves the ERK1/2 cascade and alternative promoters. Biochem Biophys Res Commun 426:172-176.

De Bruyn, T., Sempels, W., Snoeys, J., Holmstock, N., Chatterjee, S., Stieger, B., Augustijns, P., Hofkens, J., Mizuno, H., and Annaert, P. 2014. Confocal imaging with a fluorescent bile acid analogue closely mimicking hepatic taurocholate disposition. J Pharm Sci 103:1872-1881.

Degeling, M.H., Bovenberg, M.S., Tannous, M., and Tannous, B.A. Gaussia luciferase-based mycoplasma detection assay in mammalian cell culture. Methods Mol Biol 1098:47-55.

Dehghan, A., Kottgen, A., Yang, Q., Hwang, S.J., Kao, W.L., Rivadeneira, F., Boerwinkle, E., Levy, D., Hofman, A., Astor, B.C., Benjamin, E.J., van Duijn, C.M., Witteman, J.C., Coresh, J., and Fox, C.S. 2008. Association of three genetic loci with uric acid concentration and risk of gout: a genome-wide association study. Lancet 372:1953-1961.

Dong Ye, K.A.D.a.I.L. 2014. A TEM protocol for quality assurance of in vitro cellular barrier models and its application to the assessment of nanoparticle transport mechanisms across barriers. Analyst ePub Ahead of Print.

Dukes, J.D., Whitley, P., and Chalmers, A.D. 2011. The MDCK variety pack: choosing the right strain. BMC cell biology 12:43.

El-Sheikh, A.A., van den Heuvel, J.J., Koenderink, J.B., and Russel, F.G. 2008. Effect of hypouricaemic and hyperuricaemic drugs on the renal urate efflux transporter, multidrug resistance protein 4. Br J Pharmacol 155:1066-1075.

Erdman, A.R., Mangravite, L.M., Urban, T.J., Lagpacan, L.L., Castro, R.A., de la Cruz, M., Chan, W., Huang, C.C., Johns, S.J., Kawamoto, M., Stryke, D., Taylor, T.R., Carlson, E.J., Ferrin, T.E., Brett, C.M., Burchard, E.G., and Giacomini, K.M. 2006. The human organic anion transporter 3 (OAT3; SLC22A8): genetic variation and functional genomics. Am J Physiol Renal Physiol 290:F905-912.

Eric Le Ferrec, C.C., Per Artusson, David Brayden, Gèrard Fabre, Pierre Gires, François Guillou, Monique Rousset, Werner Rubas and Maria-Laura Scarino. 2001. In Vitro Models of the Intestinal Barrier. ATLA 649-668.

Feng, B., LaPerle, J.L., Chang, G., and Varma, M.V. 2010. Renal clearance in drug discovery and development: molecular descriptors, drug transporters and disease state. Expert Opin Drug Metab Toxicol 6:939-952.

Fletcher, J.I., Haber, M., Henderson, M.J., and Norris, M.D. ABC transporters in cancer: more than just drug efflux pumps. Nat Rev Cancer 10:147-156.

Folmsbee, M., Lentine, K.R., Wright, C., Haake, G., McBurnie, L., Ashtekar, D., Beck, B., Hutchison, N., Okhio-Seaman, L., Potts, B., Pawar, V., and Windsor, H. The Development of a Microbial Challenge Test with Acholeplasma laidlawii To Rate Mycoplasma-Retentive Filters by Filter Manufacturers. PDA J Pharm Sci Technol 68:281-296.

Freshney, R.I. 2010. Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications. John Wiley & Sons, Inc.

Galluzzi, L. Bravo-San Pedro, J.M. Vitale, I. Aaronson, S.A. Abrams, J.M. Adam, D. Alnemri, E.S. Altucci, L. Andrews, D. Annicchiarico-Petruzzelli, M. Baehrecke, E.H. Bazan, N.G. Bertrand, M.J. Bianchi, K. Blagosklonny, M.V. Blomgren, K. Borner, C. Bredesen, D.E. Brenner, C. Campanella, M. Candi, E. Cecconi, F. Chan, F.K. Chandel, N.S. Cheng, E.H. Chipuk, J.E. Cidlowski, J.A. Ciechanover, A. Dawson, T.M. Dawson, V.L. De Laurenzi, V. De Maria, R. Debatin, K.M. Di Daniele, N. Dixit, V.M. Dynlacht, B.D. El-Deiry, W.S. Fimia, G.M. Flavell, R.A. Fulda, S. Garrido, C. Gougeon, M.L. Green, D.R. Gronemeyer, H. Hajnoczky, G. Hardwick, J.M. Hengartner, M.O. Ichijo, H. Joseph, B. Jost, P.J. Kaufmann, T. Kepp, O. Klionsky, D.J. Knight, R.A. Kumar, S. Lemasters, J.J. Levine, B. Linkermann, A. Lipton, S.A. Lockshin, R.A. Lopez-Otin, C. Lugli, E. Madeo, F. Malorni, W. Marine, J.C. Martin, S.J. Martinou, J.C. Medema, J.P. Meier, P. Melino, S. Mizushima, N. Moll, U. Munoz-Pinedo, C. Nunez, G. Oberst, A. Panaretakis, T. Penninger, J.M. Peter, M.E. Piacentini, M. Pinton, P. Prehn, J.H. Puthalakath, H. Rabinovich, G.A. Ravichandran, K.S. Rizzuto, R. Rodrigues, C.M. Rubinsztein, D.C. Rudel, T. Shi, Y. Simon, H.U. Stockwell, B.R. Szabadkai, G. Tait, S.W. Tang, H.L. Tavernarakis, N. Tsujimoto, Y. Vanden Berghe, T. Vandenabeele, P. Villunger, A. Wagner, E.F. Walczak, H. White, E. Wood, W.G. Yuan, J. Zakeri, Z. Zhivotovsky, B. Melino, G. and Kroemer, G. 2014. Essential versus accessory aspects of cell death: recommendations of the NCCD 2015. Cell death and differentiation.

Giacomini, K.M., Huang, S.M., Tweedie, D.J., Benet, L.Z., Brouwer, K.L., Chu, X., Dahlin, A., Evers, R., Fischer, V., Hillgren, K.M., Hoffmaster, K.A., Ishikawa, T., Keppler, D., Kim, R.B., Lee, C.A., Niemi, M., Polli, J.W., Sugiyama, Y., Swaan, P.W., Ware, J.A., Wright, S.H., Yee, S.W., Zamek-Gliszczynski, M.J., and Zhang, L. Membrane transporters in drug development. Nat Rev Drug Discov 9:215-236.

Graber-Maier, A., Gutmann, H., and Drewe, J. A new intestinal cell culture model to discriminate the relative contribution of P-gp and BCRP on transport of substrates such as imatinib. Mol Pharm 7:1618-1628.

Hirouchi, M., Kusuhara, H., Onuki, R., Ogilvie, B.W., Parkinson, A., and Sugiyama, Y. 2009. Construction of triple-transfected cells [organic anion-transporting polypeptide (OATP) 1B1/multidrug resistance-associated protein (MRP) 2/MRP3 and OATP1B1/MRP2/MRP4] for analysis of the sinusoidal function of MRP3 and MRP4. Drug Metab Dispos 37:2103-2111.

Hsiang, B., Zhu, Y., Wang, Z., Wu, Y., Sasseville, V., Yang, W.P., and Kirchgessner, T.G. 1999. A novel human hepatic organic anion transporting polypeptide (OATP2). Identification of a liver-specific human organic anion transporting polypeptide and identification of rat and human hydroxymethylglutaryl-CoA reductase inhibitor transporters. J Biol Chem 274:37161-37168.

Hubatsch, I., Ragnarsson Eg Fau - Artursson, P., and Artursson, P. Determination of drug permeability and prediction of drug absorption in Caco-2 monolayers.

Ianculescu, A.G., Friesema, E.C., Visser, T.J., Giacomini, K.M., and Scanlan, T.S. 2010. Transport of thyroid hormones is selectively inhibited by 3-iodothyronamine. Mol Biosyst 6:1403-1410.

Imamura, Y., Murayama, N., Okudaira, N., Kurihara, A., Okazaki, O., Izumi, T., Inoue, K., Yuasa, H., Kusuhara, H., and Sugiyama, Y. 2011. Prediction of fluoroquinolone-induced elevation in serum creatinine levels: a case of drug-endogenous substance interaction involving the inhibition of renal secretion. Clin Pharmacol Ther 89:81-88.

Ishiguro, N., Maeda, K., Saito, A., Kishimoto, W., Matsushima, S., Ebner, T., Roth, W., Igarashi, T., and Sugiyama, Y. 2008. Establishment of a set of double transfectants coexpressing organic anion transporting polypeptide 1B3 and hepatic efflux transporters for the characterization of the hepatobiliary transport of telmisartan acylglucuronide. Drug Metab Dispos 36:796-805.

Jacob Hanna, A.H. 2009. Preservation of stem cells. Organogenesis 5:134-137.

Jonker, J.W. and Schinkel, A.H. 2004. Pharmacological and physiological functions of the polyspecific organic cation transporters: OCT1, 2, and 3 (SLC22A1-3). J Pharmacol Exp Ther 308:2-9.

Kamatani, Y., Matsuda, K., Okada, Y., Kubo, M., Hosono, N., Daigo, Y., Nakamura, Y., and Kamatani, N. Genome-wide association study of hematological and biochemical traits in a Japanese population. Nat Genet 42:210-215.

Kenzaoui, B.H., Vila, M.R., Miquel, J.M., Cengelli, F., and Juillerat-Jeanneret, L. 2012. Evaluation of uptake and transport of cationic and anionic ultrasmall iron oxide nanoparticles by human colon cells. International journal of nanomedicine 7:1275-1286.

Keppler, D. 2014. The roles of MRP2, MRP3, OATP1B1, and OATP1B3 in conjugated hyperbilirubinemia. Drug Metab Dispos 42:561-565.

Kim, K.A., Joo, H.J., and Park, J.Y. Effect of ABCG2 genotypes on the pharmacokinetics of A771726, an active metabolite of prodrug leflunomide, and association of A771726 exposure with serum uric acid level. Eur J Clin Pharmacol 67:129-134.

Kim, M.K. and Shim, C.K. 2006. The transport of organic cations in the small intestine: current knowledge and emerging concepts. Arch Pharm Res 29:605-616.

Kis, E., Ioja, E., Rajnai, Z., Jani, M., Mehn, D., Heredi-Szabo, K., and Krajcsi, P. 2011. BSEP inhibition - In vitro screens to assess cholestatic potential of drugs. Toxicol In Vitro.

Koepsell, H. 2004. Polyspecific organic cation transporters: their functions and interactions with drugs. Trends Pharmacol Sci 25:375-381.

Koepsell, H., Lips, K., and Volk, C. 2007. Polyspecific organic cation transporters: structure, function, physiological roles, and biopharmaceutical implications. Pharm Res 24:1227-1251.

Kolz, M., Johnson, T., Sanna, S., Teumer, A., Vitart, V., Perola, M., Mangino, M., Albrecht, E., Wallace, C., Farrall, M., Johansson, A., Nyholt, D.R., Aulchenko, Y., Beckmann, J.S., Bergmann, S., Bochud, M., Brown, M., Campbell, H., Connell, J., Dominiczak, A., Homuth, G., Lamina, C., McCarthy, M.I., Meitinger, T., Mooser, V., Munroe, P., Nauck, M., Peden, J., Prokisch, H., Salo, P., Salomaa, V., Samani, N.J., Schlessinger, D., Uda, M., Volker, U., Waeber, G., Waterworth, D., Wang-Sattler, R., Wright, A.F., Adamski, J., Whitfield, J.B., Gyllensten, U., Wilson, J.F., Rudan, I., Pramstaller, P., Watkins, H., Doering, A., Wichmann, H.E., Spector, T.D., Peltonen, L., Volzke, H., Nagaraja, R., Vollenweider, P., Caulfield, M., Illig, T., and Gieger, C. 2009. Meta-analysis of 28,141 individuals identifies common variants within five new loci that influence uric acid concentrations. PLoS Genet 5:e1000504.

Konig, J., Cui, Y., Nies, A.T., and Keppler, D. 2000. A novel human organic anion transporting polypeptide localized to the basolateral hepatocyte membrane. Am J Physiol Gastrointest Liver Physiol 278:G156-164.

Kopplow, K., Letschert, K., Konig, J., Walter, B., and Keppler, D. 2005. Human hepatobiliary transport of organic anions analyzed by quadruple-transfected cells. Mol Pharmacol 68:1031-1038.

Krajcsi, P. 2013. Drug-transporter interaction testing in drug discovery and
development. World J Pharmacol 9:35-46.

Kullak-Ublick, G.A., Fisch, T., Oswald, M., Hagenbuch, B., Meier, P.J., Beuers, U., and Paumgartner, G. 1998. Dehydroepiandrosterone sulfate (DHEAS): identification of a carrier protein in human liver and brain. FEBS Lett 424:173-176.

Kullak-Ublick, G.A., Ismair, M.G., Stieger, B., Landmann, L., Huber, R., Pizzagalli, F., Fattinger, K., Meier, P.J., and Hagenbuch, B. 2001. Organic anion-transporting polypeptide B (OATP-B) and its functional comparison with three other OATPs of human liver. Gastroenterology 120:525-533.

Lahoz, A., Vila, M.R., Fabre, M., Miquel, J.M., Rivas, M., Maines, J., Castell, J.V., and Gomez-Lechon, M.J. 2013. An in vitro tool to assess cytochrome P450 drug biotransformation-dependent cytotoxicity in engineered HepG2 cells generated by using adenoviral vectors. Toxicology in vitro : an international journal published in association with BIBRA 27:1410-1415.

Lee, W., Belkhiri, A., Lockhart, A.C., Merchant, N., Glaeser, H., Harris, E.I., Washington, M.K., Brunt, E.M., Zaika, A., Kim, R.B., and El-Rifai, W. 2008. Overexpression of OATP1B3 confers apoptotic resistance in colon cancer. Cancer Res 68:10315-10323.

Li, A.P. 2005. Preclinical in vitro screening assays for drug-like properties. Drug discovery today. Technologies 2:179-185.

Lufino, M.M., Edser, P.A., and Wade-Martins, R. 2008. Advances in high-capacity extrachromosomal vector technology: episomal maintenance, vector delivery, and transgene expression. Mol Ther 16:1525-1538.

Maddox, C.B., Rasmussen, L., and White, E.L. 2008. Adapting Cell-Based Assays to the High Throughput Screening Platform: Problems Encountered and Lessons Learned. JALA (Charlottesville, Va.) 13:168-173.

Maher, J.M., Dieter, M.Z., Aleksunes, L.M., Slitt, A.L., Guo, G., Tanaka, Y., Scheffer, G.L., Chan, J.Y., Manautou, J.E., Chen, Y., Dalton, T.P., Yamamoto, M., and Klaassen, C.D. 2007. Oxidative and electrophilic stress induces multidrug resistance-associated protein transporters via the nuclear factor-E2-related factor-2 transcriptional pathway. Hepatology 46:1597-1610.

Maria S. Balda, J.A.W., Catalina Flores, Sirenia González, Marcelino Cereijido, and Karl Matter. 1996. Functional Dissociation of Paracellular Permeability and Transepithelial Electrical Resistance and Disruption of the Apical-Basolateral Intramembrane Diffusion Barrier by Expression of a Mutant Tight Junction Membrane Protein The Journal of Cell Biology 134:1031-1049.

Markowska, M., Oberle R Fau - Juzwin, S., Juzwin S Fau - Hsu, C.P., Hsu Cp Fau - Gryszkiewicz, M., Gryszkiewicz M Fau - Streeter, A.J., and Streeter, A.J. Optimizing Caco-2 cell monolayers to increase throughput in drug intestinal absorption analysis.

Meunier V, B.M., Julian B, Marti E, Guillou F, Berger Y, Fabre G. 2000. Expression and induction of CYP1A1/1A2, CYP2A6 and CYP3A4 in primary cultures of human hepatocytes: a 10-year follow-up. Xenobiotica 30:589-607.

Mita, S., Suzuki, H., Akita, H., Hayashi, H., Onuki, R., Hofmann, A.F., and Sugiyama, Y. 2006. Vectorial transport of unconjugated and conjugated bile salts by monolayers of LLC-PK1 cells doubly transfected with human NTCP and BSEP or with rat Ntcp and Bsep. Am J Physiol Gastrointest Liver Physiol 290:G550-556.

Mita, S., Suzuki, H., Akita, H., Stieger, B., Meier, P.J., Hofmann, A.F., and Sugiyama, Y. 2005. Vectorial transport of bile salts across MDCK cells expressing both rat Na+-taurocholate cotransporting polypeptide and rat bile salt export pump. Am J Physiol Gastrointest Liver Physiol 288:G159-167.

Morgan, R.E., Trauner, M., van Staden, C.J., Lee, P.H., Ramachandran, B., Eschenberg, M., Afshari, C.A., Qualls, C.W., Jr., Lightfoot-Dunn, R., and Hamadeh, H.K. 2010. Interference with bile salt export pump function is a susceptibility factor for human liver injury in drug development. Toxicol Sci 118:485-500.

Motohashi, H. and Inui, K. 2013. Multidrug and toxin extrusion family SLC47: physiological, pharmacokinetic and toxicokinetic importance of MATE1 and MATE2-K. Mol Aspects Med 34:661-668.

Motohashi, H., Sakurai, Y., Saito, H., Masuda, S., Urakami, Y., Goto, M., Fukatsu, A., Ogawa, O., and Inui, K. 2002. Gene expression levels and immunolocalization of organic ion transporters in the human kidney. J Am Soc Nephrol 13:866-874.

Nakamura, T., Yonezawa, A., Hashimoto, S., Katsura, T., and Inui, K. 2010. Disruption of multidrug and toxin extrusion MATE1 potentiates cisplatin-induced nephrotoxicity. Biochem Pharmacol 80:1762-1767.

Obaidat, A., Roth, M., and Hagenbuch, B. 2012. The expression and function of organic anion transporting polypeptides in normal tissues and in cancer. Annu Rev Pharmacol Toxicol 52:135-151.

Olarerin-George, O. and Hogenesch, J.B. 2014. Assessing the prevalence of mycoplasma contamination in cell culture via a survey of
NCBI’s RNA-seq archive. bioRxiv.

Oswald, S., Groer, C., Drozdzik, M., and Siegmund, W. 2013. Mass spectrometry-based targeted proteomics as a tool to elucidate the expression and function of intestinal drug transporters. AAPS J 15:1128-1140.

Pratt, E., Sissung, T.M., and Figg, W.D. 2012. Loss of OATP1B3 function causes Rotor syndrome: implications for potential use of inhibitors in cancer. Cancer Biol Ther 13:1374-1375.

Sampson, K.E., Brinker, A., Pratt, J., Venkatraman, N., Xiao, Y., Blasberg, J., Steiner, T., Bourner, M., and Thompson, D.C. Zinc Finger Nuclease-mediated Gene Knockout Results in Loss of Transport Activity for P-glycoprotein, BCRP, and MRP2 in Caco-2 Cells. Drug Metab Dispos.

Sekine, T., Cha, S.H., and Endou, H. 2000. The multispecific organic anion transporter (OAT) family. Pflugers Arch 440:337-350.

Shu, Y., Brown, C., Castro, R.A., Shi, R.J., Lin, E.T., Owen, R.P., Sheardown, S.A., Yue, L., Burchard, E.G., Brett, C.M., and Giacomini, K.M. 2008. Effect of genetic variation in the organic cation transporter 1, OCT1, on metformin pharmacokinetics. Clin Pharmacol Ther 83:273-280.

Sigel, E. 1990. Use of Xenopus oocytes for the functional expression of plasma membrane proteins. J Membr Biol 117:201-221.

Stacey, G.N. 1997. Standardization in animal cell technology. Folia microbiologica 42:113-116.

Staud, F., Cerveny, L., Ahmadimoghaddam, D., and Ceckova, M. 2013. Multidrug and toxin extrusion proteins (MATE/SLC47); role in pharmacokinetics. Int J Biochem Cell Biol 45:2007-2011.

Stieger, B. and Beuers, U. 2011. The canalicular bile salt export pump BSEP (ABCB11) as a potential therapeutic target. Curr Drug Targets 12:661-670.

Tamai, I., Nezu, J., Uchino, H., Sai, Y., Oku, A., Shimane, M., and Tsuji, A. 2000. Molecular identification and characterization of novel members of the human organic anion transporter (OATP) family. Biochem Biophys Res Commun 273:251-260.

Uchiumi, T., Kohno, K., Tanimura, H., Hidaka, K., Asakuno, K., Abe, H., Uchida, Y., and Kuwano, M. 1993. Involvement of protein kinase in environmental stress-induced activation of human multidrug resistance 1 (MDR1) gene promoter. FEBS Lett 326:11-16.

van de Steeg, E., Stranecky, V., Hartmannova, H., Noskova, L., Hrebicek, M., Wagenaar, E., van Esch, A., de Waart, D.R., Oude Elferink, R.P., Kenworthy, K.E., Sticova, E., al-Edreesi, M., Knisely, A.S., Kmoch, S., Jirsa, M., and Schinkel, A.H. 2012. Complete OATP1B1 and OATP1B3 deficiency causes human Rotor syndrome by interrupting conjugated bilirubin reuptake into the liver. J Clin Invest 122:519-528.

van Herwaarden, A.E. and Schinkel, A.H. 2006. The function of breast cancer resistance protein in epithelial barriers, stem cells and milk secretion of drugs and xenotoxins. Trends Pharmacol Sci 27:10-16.

Volokhov, D.V., Graham, L.J., Brorson, K.A., and Chizhikov, V.E. Mycoplasma testing of cell substrates and biologics: Review of alternative non-microbiological techniques. Mol Cell Probes 25:69-77.

Weksler, B., Romero Ia Fau - Couraud, P.-O., and Couraud, P.O. 2013. The hCMEC/D3 cell line as a model of the human blood brain barrier.

West, K.L., Ramjiganesh, T., Roy, S., Keller, B.T., and Fernandez, M.L. 2002. 1-[4-[4[(4R,5R)-3,3-Dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-di oxido-1-benzothiepin-5-yl]phenoxy]butyl]-4-aza-1-azoniabicyclo[2.2.2]octane methanesulfonate (SC-435), an ileal apical sodium-codependent bile acid transporter inhibitor alters hepatic cholesterol metabolism and lowers plasma low-density lipoprotein-cholesterol concentrations in guinea pigs. J Pharmacol Exp Ther 303:293-299.

Wils, P., Warnery A Fau - Phung-Ba, V., Phung-Ba V Fau - Scherman, D., and Scherman, D. 1994. Differentiated intestinal epithelial cell lines as in vitro models for predicting the intestinal absorption of drugs.

Woodward, O.M., Kottgen, A., Coresh, J., Boerwinkle, E., Guggino, W.B., and Kottgen, M. 2009. Identification of a urate transporter, ABCG2, with a common functional polymorphism causing gout. Proc Natl Acad Sci U S A 106:10338-10342.

Xia, C.Q., Milton, M.N., and Gan, L.S. 2007. Evaluation of drug-transporter interactions using in vitro and in vivo models. Curr Drug Metab 8:341-363.

Xiang, X., Han, Y., Neuvonen, M., Pasanen, M.K., Kalliokoski, A., Backman, J.T., Laitila, J., Neuvonen, P.J., and Niemi, M. 2009. Effect of SLCO1B1 polymorphism on the plasma concentrations of bile acids and bile acid synthesis marker in humans. Pharmacogenet Genomics 19:447-457.

Yeh, G.C., Lopaczynska, J., Poore, C.M., and Phang, J.M. 1992. A new functional role for P-glycoprotein: efflux pump for benzo(alpha)pyrene in human breast cancer MCF-7 cells. Cancer Res 52:6692-6695.

Yokoo, S., Yonezawa, A., Masuda, S., Fukatsu, A., Katsura, T., and Inui, K. 2007. Differential contribution of organic cation transporters, OCT2 and MATE1, in platinum agent-induced nephrotoxicity. Biochem Pharmacol 74:477-487.

Yonezawa, A. and Inui, K. 2011. Organic cation transporter OCT/SLC22A and H(+)/organic cation antiporter MATE/SLC47A are key molecules for nephrotoxicity of platinum agents. Biochem Pharmacol 81:563-568.

Young, L., Sung, J., Stacey, G., and Masters, J.R. Detection of Mycoplasma in cell cultures. Nat Protoc 5:929-934.

Yu, M., Ocana, A., and Tannock, I.F. 2013. Reversal of ATP-binding cassette drug transporter activity to modulate chemoresistance: why has it failed to provide clinical benefit? Cancer Metastasis Rev 32:211-227.