Stokes William S, Casati Silvia, Strickland Judy, Paris Michael
The National Toxicology Program Interagency Center for the Evaluation of Alternative Toxicological Methods, Research Triangle Park, North Carolina, USA.
Curr Protoc Toxicol. 2008 May;Chapter 20:Unit 20.4. doi: 10.1002/0471140856.tx2004s36.
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.
体外细胞毒性试验可用作替代毒性试验，以减少急性经口毒性试验所需的动物总数。本单元介绍了两种使用中性红摄取法（NRU）测定受试物体外细胞毒性的方法，以及如何利用体外数据确定体内急性经口全身毒性试验（如上下法或急性毒性分级法）的起始剂量。使用NRU方法确定急性经口毒性试验的起始剂量可能会减少所需动物的数量，对于毒性相对较大的物质，这种方法还可能减少因严重毒性而死亡或需要实施安乐死的动物数量。一项实验室间验证研究表明，这些方法对于上述目的是有用且可重复的。两个标准化方案提供了用啮齿动物和人类细胞进行NRU试验的详细操作说明。
