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KMID : 1177720100040010005
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Journal of Alternatives to Animal Experiments
2010 Volume.4 No. 1 p.5 ~ p.9
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In vivo Micronucleus Assay in Peripheral Blood by Flow Cytometric Analysis
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Kim Joo-Hwan
Yum Young-Na
Kim Hee-Yun
Kim Kyong-A
Kim Ji-Eun
Park Sue-Nie
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Abstract
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The micronucleus assay is the most widely utilized in vivo system for evaluating chemicals potential to induce chromosome breaks or to poison mitotic spindle apparatus. The bone marrow erythrocyte micronucleus assay is widely used for regulatory studies conducted to evaluate the potential for induction of chromosomal damage or chromosome loss due to exposure to drugs, food additives, pesticides, industrial chemicals, and other products. Peripheral blood reticulocytes(RET) have been accepted as an appropriate target for micronucleus (MN) assessment for both acute and cumulative damage. However, there is a problem when using rat peripheral blood reticulocytes, since the rat spleen selectively removes MN?RET from the circulation. The development of automated flow cytometric(FCM), anti?CD71?based methods for evaluating micronucleus frequencies in reticulocytes has great potential for improving the sensitivity, reproducibility, and throughput of the traditional in vivo rodent MN assay that uses microscopy?based methods. There were some validation studies of the FCM evaluation methods in many species. For instance it has been demonstrated that for mice, rats, dogs, monkey, and humans. Although spleen?dependent reduction in circulating MN?RET frequency resulting from genotoxic exposures attenuates the magnitude of MN?RET frequencies in peripheral blood relative to that in bone marrow, the analytical and statistical advantages of the FCM measurement in peripheral blood more than offset
the higher frequencies observed for bone marrow smears analyzed. In addition to the analytical advantages of the
FCM method, the ready accessibility of the small blood samples required for the FCM assay make possible the
integration of the MN frequency assessment with routine toxicology studies. It is not necessary to conduct separate in vivo studies for the purpose of evaluating the potential for chromosomal damage in rodent bone marrow but that more reliable information can be obtained by FCM analysis of peripheral blood samples obtained from rodent studies during the course of routine toxicological evaluations.
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KEYWORD
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In vivoMicronucleus, Flow Cytometric Analysis, Peripheral Blood
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