FLOW CYTOMETRIC RAT MICRONUCLEUS ASSAY

流式细胞术大鼠微核测定

• 批准号: 6382267
• 负责人: DOROTHEA K TOROUS
• 金额: $ 27.66万
• 依托单位: LITRON LABORATORIES, LTD.
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2002-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6382267
• 关键词: DNA damage; bioassay; biomedical automation; blood tests; bone marrow; cytotoxicity; flow cytometry; laboratory rat; method development; micronucleus; stainings; toxicant screening; toxin

Chemicals play an important role in our quality of life, thus it is important to characterize the toxicity of new chemicals so that harmful agents can be highlighted prior to public exposure. The micronucleus (MN) assay is the most widely used system or evaluating a chemical's potential to cause chromosomal damage. This research project will broaden the database of genotoxins analyzed by flow cytometry (FCM), assessing whether rat peripheral blood (after acute or subchronic treatment) is an appropriate compartment for detecting damage caused by genotoxic compounds. Since rat bone marrow is currently accepted by regulatory agencies worldwide, a major line of investigation is to further develop reliable methods for preparing bone marrow samples for FCM analysis. A significant in te rest for automating the rat peripheral blood assay is that the spleen of this species behaves similarly to that of humans. Therefore, this work is extremely important towards our long-term goal of analyzing human blood for the presence of MN. When we are able to measure MN in human blood, it might be possible to determine the genotoxic effects of accidental human exposure and even biomonitor patients receiving chemotherapeutic treatments. PROPOSED COMMERCIAL APPLICATION: Litron, together with the Stratagene Corporation, is developing micronucleus kits for rat peripheral blood and bone marrow. The kits will be in two formats: the first allows researchers to send blood samples to Litron for MN analysis (microFlow kits)to analyze for MN, and the second allows researchers to analyze for MN in-house (micro FlowPlus kits). The pharmaceutical community has responded enthusiastically and has asked for the development of rat kits specifically for their in-house use in addition to the kits, we expect our genetic toxicology contract testing to increase as this advanced technology is refined and brought to the marketplace.

化学品在我们的生活质量中发挥着重要作用，因此，重要的是要描述新化学品的毒性，以便在公众接触之前突出有害物质。微核（MN）试验是最广泛使用的系统或评估化学品引起染色体损伤的潜力。本研究项目将扩大通过流式细胞术（FCM）分析的遗传毒素数据库，评估大鼠外周血（急性或亚慢性治疗后）是否是检测遗传毒性化合物引起的损伤的适当隔室。由于大鼠骨髓目前已被全球监管机构接受，因此研究的主要方向是进一步开发用于FCM分析的骨髓样品制备的可靠方法。大鼠外周血分析自动化的一个重要目的是，该物种的脾脏与人类的脾脏表现相似。因此，这项工作对于我们分析人类血液中MN存在的长期目标非常重要。当我们能够测量人类血液中的MN时，就有可能确定意外人体暴露甚至接受化疗的生物监测患者的遗传毒性效应。拟定商业应用：Litron与Stratagene公司正在开发用于大鼠外周血和骨髓的微核试剂盒。这些试剂盒将有两种形式：第一种允许研究人员将血液样本发送到Litron进行MN分析（microFlow试剂盒）以分析MN，第二种允许研究人员在内部分析MN（microFlowPlus试剂盒）。制药界对此反应热烈，并要求开发专门用于内部使用的大鼠试剂盒。除了试剂盒外，我们预计，随着这项先进技术的完善和推向市场，我们的遗传毒理学合同测试将增加。

项目成果

Comparative scoring of micronucleated reticulocytes in rat peripheral blood by flow cytometry and microscopy.

通过流式细胞术和显微镜对大鼠外周血中微核网织红细胞的比较评分。

• DOI: 10.1093/toxsci/kfg143
• 发表时间: 2003
• 期刊: Toxicological sciences : an official journal of the Society of Toxicology.
• 作者: Torous,DorotheaK;Hall,NikkiE;Murante,FrancisG;Gleason,SarahE;Tometsko,CarolR;Dertinger,StephenD
• 通讯作者: Dertinger,StephenD