Neutral red uptake cytotoxicity tests for estimating starting doses for acute oral toxicity tests.
In vitro cytotoxicity assays can be used as alternative toxicity tests to reduce the total number of animals needed for acute oral toxicity tests. This unit describes two methods for determining the in vitro cytotoxicity of test substances using neutral red uptake (NRU) and using the in vitro data to determine starting doses for in vivo acute oral systemic toxicity tests, e.g., the up-and-down procedure or the acute toxic class method. The use of the NRU methods to determine starting doses for acute oral toxicity tests may reduce the number of animals required, and for relatively toxic substances, this approach may also reduce the number of animals that die or require humane euthanasia due to severe toxicity. An interlaboratory validation study has demonstrated that the methods are useful and reproducible for these purposes. Two standardized protocols provide details for performing NRU tests with rodent and human cells.
Stokes
William S
WS
The National Toxicology Program Interagency Center for the Evaluation of Alternative Toxicological Methods, Research Triangle Park, North Carolina, USA.
Casati
Silvia
S
Strickland
Judy
J
Paris
Michael
M
D000826:Animal Testing Alternatives; D000818:Animals; D041702:BALB 3T3 Cells; D001692:Biological Transport / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism*; D006801:Humans; D007202:Indicators and Reagents / Q000008:administration & dosage / Q000493:pharmacokinetics*; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism*; D051379:Mice; D009499:Neutral Red / Q000008:administration & dosage / Q000493:pharmacokinetics*; D009994:Osmolar Concentration; D012995:Solubility; D018675:Toxicity Tests
D003470:Culture Media; D007202:Indicators and Reagents; D009499:Neutral Red