FLOW CYTOMETRIC RAT MICRONUCLEUS ASSAY

微核（MN）试验是最广泛使用的体内试验 遗传毒性试验，并建议用于新的 化学品和配方的监管机构在世界各地。的目的 这项研究计划是评估是否修改我们的 经验证的基于流式细胞术（FCM）的小鼠MN测定可用于 准确测量骨髓和外周血中的MN频率 老鼠。大鼠骨髓试验目前使用 传统的显微镜方法，成本高，繁琐，费时， 消耗。 由于大鼠是毒理学中使用最广泛的啮齿动物， 在药代动力学研究中，大鼠外周血测试系统将是 对于整合体内终点非常有用， 减少动物需求。如果一个敏感的微核试验可以 纳入其他大鼠研究，关于 潜在的遗传毒性将在不使用额外的 动物或其他试验材料。一种自动检测方法， 在大鼠中的致染色体断裂活性将代表一个显著的进步， 现有的方法在周转时间、成本和可靠性方面存在缺陷。 实验计划：1）确定准确分析的方法 用流式细胞术检测大鼠外周血和骨髓MN的变化; 2）评价 测量时亚慢性给药和出血方案的范围 外周血MN以及将MN终点与 这些研究。通过使用FCM结合适当的固定和 染色程序，我们希望开发一种可靠的方法来计数MN 在大鼠外周血和骨髓中。 这些测量的自动化将使Litron能够提供另一种 技术创新和改进的遗传毒理学服务。这些 测试系统将提供比现有协议更大的优势 通过提高灵敏度和效率，同时 减少用于遗传毒理学测试的动物数量。这 该方法提供了巨大的商业潜力，因为工业将节省资金， 动物、试验材料和时间将MN分析外包给专家 像Litron这样的设施。 拟议的商业应用： 外周血微核的自动化检测方法 大鼠骨髓将代表着比现有的 方法在周转时间，成本和可靠性方面。拟议 研究计划将进一步建立Litron实验室作为一个 专家测试设施，为工业界、学术界和政府提供 具有高度可靠的遗传毒性数据。我们计划开发一个研究工具包 其中包括研究人员获取血液所需的试剂 用于Litron后续MN分析的样品。

The micronucleus (MN) assay is the most widely utilized in vivo genotoxicity assay and is recommended for the safeLy evaluation of new chemicals and formulations by regulatory agencies worldwide. The aim of this research program is to evaluate whether modifications to our validated flow cytometry (FCM) based mouse MN assay can be used to accurately measure MN frequencies in the hone marrow and peripheral blood of rats. The rat bone marrow assay is currently performed using traditional microscopic methods, which are costly, tedious and time- consuming. Since rats are the most widely used rodent in toxicology and pharmacokinetic studies, a rat peripheral blood test system would be extremely useful for integrating in vivo endpoints and for significantly decreasing animal requirements. If a sensitive micronucleus test could be integrated into other rat studies, preliminary information regarding potential genotoxicity would be obtained without the use of additional animals or additional test material. An automated method of detecting clastogenic activity in rats would represent a significant advance over existing methods in terms of turnaround-time, cost and reliability. Experiments are planned to:l) determine methods for the accurate analysis of rat peripheral blood and bone marrow MN by FCM; and 2) evaluate the latitude of subchronic dosing and bleeding regimens when measuring peripheral blood MN and the ability to integrate the MN endpoint with these studies. By utilizing FCM in conjunction with proper fixing and staining procedures we hope to develop a reliable method to enumerate MN in rat peripheral blood and hone marrow. Automation of these measurements would enable Litron to offer another technologically innovative and improved genetic toxicology service. These test systems will offer significant advantages over existing protocols by increasing the sensitivity and efficiency, while simultaneously decreasing the number of animals used in genetic toxicology testing. This method offers enormous commercial potential as industry would save money, animals, test material and time contracting out MN analyses to an expert facility such as Litron. PROPOSED COMMERCIAL APPLICATIONS: An automated method of detecting micronuclei in the peripheral blood and bone marrow of rats would represent a significant advance over existing methods in terms of turn-around-time, cost and reliability. The proposed research initiative will further establish Litron Laboratories as an expert testing facility which provides industry, academia and government with highly reliable genotoxicity data. We plan to develop a research kit which would include reagents necessary for researchers to obtain blood samples for subsequent MN analyses by Litron.