Cryopreserved human hepatocytes

Shitara, Y., Lu, C., Li, A. P., Y. Kato, Y., Suzuki, H., Ito, K., Itoh, T., Sugiyama, Y., Cryopreserved human hepatocytes as a tool for the prediction of in vivo transport and transporter-mediated drug-drug interactions, Abstract of International Conference on Drug Interaction, Hamamatsu, October 21-23, 1999, p. 87. 

Shitara Y, Li AP, Kato Y, et al. Function of uptake transporters for taurocholate and estradiol 17b - D-glucuronide in cryopreserved human hepatocytes. Dmg Metab Pharmacokinet 2003 18 33 41. 

Zhao P, Kunze KL, Lee CA. Evaluation of time-dependent inactivation of CYP3A in cryopreserved human hepatocytes. Drug Metab Dispos 2005 33(6) 853-861. 

Niro, R., Byers, J., Fournier, R., and Bachmann, K., Application of a convective-dispersion model to predict in vivo hepatic clearance from in vitro measurements utilizing cryopreserved human hepatocytes, Current Drug Metabolism, Vol. 4, No. 5, 2003, pp. 357-369. 

Lee SH, Slattery JT (1997) Cytochrome P450 isozymes involved in lisofylline metabolism to pentoxifylline in human liver microsomes. Drug Metab Dispos 25 1354-1358 Li AP (1999) Cryopreserved human hepatocytes characterization of DME activities and applications in higher throughput screening assays for hepatotoxicity, metabolic stability and drug-drug interaction potential. Chem Biol Interact 121 17-35... 

Cryopreserved human hepatocytes Cryopreserved human hepatocytes are extremely useful for the evaluation of drug metabolism, but in general cannot be cultured due to their impaired cell-attachment. There are preparations of cryopreserved human hepatocytes with high plating efficiency and therefore can be cultured for induction studies. It has been shown that hepatocytes cultured after cryopreserva-tion are responsive to CYP1A and 3A inducers, but they have a significantly lower basal (uninduced) levels of these enzymes. Cryopreserved human hepatocytes therefore represent a more convenient experimental model than freshly isolated human hepatocytes for enzyme induction studies (Roymans etal. 2005). 

Roymans D, Van Looveren C, Leone A et al. (2004) Determination of cytochrome P450 1A2 and cytochrome P450 3A4 induction in cryopreserved human hepatocytes. Biochem Pharmacol 67 427 137... 

Hepatotoxicity—enzyme release and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazodium bromide (MTT) assay on fresh or cryopreserved (human) hepatocytes Endocrine disruption—yeast and mammalian reporter gene assay (oestrogenicity and adrogenicity)... 

In vitro system Cryopreserved human hepatocytes pooled from two donors (male, female)... 

Day 0 Plate human hepatocytes (freshly isolated or plateable cryopreserved human hepatocytes)... 

Bi YA, Kazolias D, Duignan DB (2006) Use of cryopreserved human hepatocytes in sandwich culture to measure hepatobiliary transport. Drug Metab Dispos Biol Fate Chem 34 1658-1665... 

Zanelli U, Caradonna NP, Hallifax D et al (2012) Comparison of cryopreserved HepaRG cells with cryopreserved human hepatocytes for prediction of clearance for 26 drugs. Drug Metab Dispos 40 104-110... 

Jouin, D. et al., Cryopreserved human hepatocytes in suspension are a convenient high throughput tool for the prediction of metabolic clearance, Eur. J. Pharm. Biopharm., 63, 347, 2006. 

Hewitt NJ, Lloyd S, Haydan M, Butler R, Sakai Y, Springer R, Fackett A, and Li AP (2002) Correlation between troglitazone cytotoxicity and drug metabolic enzyme activities in cryopreserved human hepatocytes. Chem Biol Interact 142 73-82 Holcik M, Sonenberg N (2005) Translational control in stress and apoptosis. Nature Rev Mol Cell Biol 6 318-327... 

Hallifax D, Rawden HC, Hakooz N, Houston JB. Prediction of metabolic clearance using cryopreserved human hepatocytes kinetic characteristics for five benzodiazepines. Drug Metab Dispos 2005 33 1852-1858. 

Cryopreserved human hepatocytes from three male and two female donors or freshly isolated male rat hepatocytes are analyzed for viabilities (75-85%) using the trypan blue exclusion methods. Incubations are performed by suspending the hepatocytes in Krebs-biocarbonate buffer followed by addition of a H-labeled compound in methanol. The specific radioactivity of the compounds is 100 Ci/ mol. The final concentration of test compound in the suspension is 10 xM in a final volume of 1 mL (1x10 cells/mL), and the final concentration of methanol does not exceed 0.2% (v/v). Incubations are allowed to proceed at 37 °C for 1 h, and are quenched with acetonitrile (5 mL). The remaining procedures are the same as described in Section 14.2.2 (the protocol for in vitro covalent protein binding in human or rat liver microsomes a test-tube method). Covalent protein binding values in pmol-equiv./mg protein are estimated based on the residual radioactivity in the protein pellets. 

Kafert-Kasting S, Alexandrova K, Barthold M, Laube B, Friedrieh G, Arseniev L, Hengstler JG. Enzyme induetion in cryopreserved human hepatocyte eultures. Toxicology 2006 220 117-125. 

Reinach B, de Sousa G, Dostert P, Ings R, Gugenheim J, Rahmani R. Comparative effects of rifabutin and rifampicin on cytochromes P450 and UDP-glucuronosyl-transferases expression in fresh and cryopreserved human hepatocytes. Chem Biol Interact 1999 121 37-48. 

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