In a pilot study, we tested 20 randomly-selected chemicals for their cytotoxicity toward the HPCT-1E3 cell model, in order to prove the ability of this in vitro model to predict human acute in vivo toxicity. The study revealed that, in contrast to most other in vitro models, results from the HPCT-1E3 cell-based system show better correlation with the more-relevant human acute lethal doses, whereas results from most other systems have a high predictivity for human lethal serum concentrations. For the prevalidation of the HPCT-1E3 model as a surrogate for regulatory acute in vivo toxicity tests, we have now expanded the list of tested chemicals to 57 substances, and have compared the results with data from the HepG2 cell assay. Again, a better correlation of HPCT-1E3 IC50 values with human oral lethal doses, as compared to correlation with human lethal serum concentrations, was observed after the pooling of all the tested substances (r2 = 0.53 [P < 0.001] and r2 = 0.41 [P = 0.009], respectively). Therefore, the HPCT-1E3 in vitro model may be a valuable tool for prediction of human oral toxicity, and may help to further reduce the number of animals used for in vivo toxicity tests.
RussellW.M.S. & BurchR.L. (1959). The Principles of Humane Experimental Technique, 238pp. London, UK: Methuen.
2.
BondessonI., EkwallB., HellbergS., RomertL., StenbergK. & WalumE. (1989). MEIC — a new international multicenter project to evaluate the relevance to human toxicity of in vitro cytotoxicity tests. Cell Biology & Toxicology5, 331–347.
3.
EkwallB. (1999). Overview of the final MEIC results: II. The in vitro–in vivo evaluation, including the selection of a practical battery of cell tests for prediction of acute lethal blood concentrations in humans. Toxicology in Vitro13, 665–673.
4.
PetzingerE., FöllmannW., BlumrichM., WaltherP., HentschelJ., BetteP., MauriceM. & FeldmannG. (1994). Immortalization of rat hepatocytes by fusion with hepatoma cells. I. Cloning of a hepatocytoma cell line with bile canaliculi. European Journal of Cell Biology64, 328–338.
5.
UteschD., ArandM., ThomasH., PetzingerE. & OeschF. (1992). Xenobiotic-metabolizing enzyme activities in hybrid cell lines established by fusion of primary rat liver parenchymal cells with hepatoma cells. Xenobiotica22, 1451–1457.
6.
HalwachsS., KneuerC. & HonschaW. (2005). Endogenous expression of liver-specific drug transporters for organic anions in the rat hepatocytoma fusion cell line HPCT-1E3. European Journal of Cell Biology84, 677–686.
7.
KneuerC., LakomaC. & HonschaW. (2007). Prediction of acute toxicity in HPCT-1E3 hepatocytoma cells with liver-like transport activities. ATLA35, 411–420.
8.
HonschaW. & PetzingerE. (1999). Characterization of the bile acid sensitive methotrexate carrier of rat liver cells. Naunyn-Schmiedeberg's Archives of Pharmacology359, 411–419.
9.
CuiY., KönigJ., NiesA.T., PfannschmidtM., HergtM., FrankeW.W., AltW., MollR. & KepplerD. (2003). Detection of the human organic anion transporters SLC21A6 (OATP2) and SLC21A8 (OATP8) in liver and hepatocellular carcinoma. Laboratory Investigation83, 527–538.
10.
Anon. (2000). The Multicenter Evaluation of In Vitro Cytotoxicity (MEIC) Summary, 66pp. Research Triangle Park, NC, USA: National Toxicology Program (NTP) Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM).
11.
LorgueG., LechenetJ. & RiviereA. (1996). Specific poisons. In Clinical Veterinary Toxicology (ed. LorgueG., LechenetJ. & RiviereA.), pp. 31–195. Oxford, UK: Blackwell Science.
12.
WalumE. (1998). Acute oral toxicity. Environmental Health Perspectives106, 497–503.
13.
SchrenkD., KnerrS. & SchmitzH.J. (2008). Monographien. In Giftlexikon (ed. SeegerR. & NeumannG.), 462pp. Stuttgart, Germany: Deutscher Apotheker Verlag.
14.
AlthausF.R. (undated). CliniPharm. Zurich, Switzerland: Institut für Veterinärpharmakologie und -toxikologie. Available at: http://www.vetpharm.uzh.ch/perldocs/wirksto.htm. (Accessed 03.10.13).
FriedmanS.J. & SkehanP. (1980). Membrane-active drugs potentiate the killing of tumor cells by D-glucosamine. Proceedings of the National Academy of Sciences of the USA77, 1172–1176.
18.
PaillardF., FinotF., MoucheI., PrenezA. & VericatJ.A. (1999). Use of primary cultures of rat hepatocytes to predict toxicity in the early development of new chemical entities. Toxicology in Vitro13, 693–700.
19.
ClothierR., Gómez-LechónM.J., Kinsner-OvaskainenA., Kopp-SchneiderA., O'ConnorJ.E., PrietoP. & StanzelS. (2012). Comparative analysis of eight cytotoxicity assays evaluated within the ACuteTox Project. Toxicology in Vitro27, 1347–1356.
20.
GarleM.J., FentemJ.H. & FryJ.R. (1994). In vitro cytotoxicity tests for the prediction of acute toxicity in vivo. Toxicology in Vitro8, 1303–1312.
HalleW. (2003). The Registry of Cytotoxicity: Toxicity testing in cell cultures to predict acute toxicity (LD50) and to reduce testing in animals. ATLA31, 189–198.
23.
HalleW., LiebschM., TraueD. & SpielmannH. (1997). Reduction of the numbers of animals used for the classification of the acute oral toxicity of chemicals by taking into account cytotoxicity data from the Registry of Cytotoxicity. ALTEX14, 8–15.
24.
SchledeE., MischkeU., RollR. & KayserD. (1992). A national validation study of the acute-toxic-class method — an alternative to the LD50 test. Archives of Toxicology66, 455–470.
EURL ECVAM (2013). Recommendation on the 3T3 NRU Assay for Supporting the Identification of Substances Not Requiring Classification for Acute Oral Toxicity: JRC Scientific and Policy Reports, 46pp. Ispra, Italy: European Commission, Joint Research Centre, Institute for Health and Consumer Protection (IHCP). Available at: http://ihcp.jrc.ec.europa.eulour/absleur/-ecvamleur/-ecvam-recommendationslfiles-3t3/ReqNoJRC79556lbna25946enn.pdf (Accessed 28.08.13).
