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Three Dimensional Nucleus Imaging analysis & in vitro Genotoxicity Test through Advanced Cytokinesis-Block Micronucleus Assay

  • Journal of Alternatives to Animal Experiments
  • Abbr : J Altern Anim Exp
  • 2018, 12(1), pp.1-7
  • DOI : 10.23032/jaae.2018.12.1.001
  • Publisher : Korean Society for Alternatives to Animal Experiments
  • Research Area : Medicine and Pharmacy > Veterinary > Basic Veterinary Medicine > Laboratory Animal Medicine
  • Received : December 18, 2018
  • Accepted : December 27, 2018
  • Published : December 29, 2018

Hye-Ji Kim 1 In-Cheol Seo 2 Whui-Su Shim 3 Chae-Eun Lee 4 Won-Seok Chang 5 Hyun-Su Lee 6 Sun-Hyun Park 1

1한국화학연구원 부설 안전성평가연구소 차세대의약평가연구단
2합천노인병원
3한국과학기술원 화학과
4한국화학연구원 부설 안전성평가연구소 유전독성연구그룹
5을지대학교 의과대학
6계명대학교 의과대학

ABSTRACT

The cytokinesis-block micronucleus cytome assay is a measuring of DNA damage in once-divided binucleated (BN) cells scoring micronuclei (MNi) that are widely used as biomarkers of cancer risk in humans on drugs or chemicals. However, this assay is still challenging due to the speed of sample processing and the toxicity of organic chemicals on researcher. To address these problems, we developed a new and convenient method for the micronucleus cytome assay. This method by DAPI stained micronucleus detection is non-toxic to the researcher and is being simplified because it does not require the use of methanol and acetic acid for fixing cell. Especially, three dimensional imaging of nucleus by formalin fixed DAPI staining provides more detailed information for testing the effects of chemical and bio-materials, compared to the classic method. In this study, we examine the spatial distribution of the TANNylated GFPs by advanced cytokinesis-block micronucleus (ACBMN) assay. The TANNylated GFPs were precisely observed in the inner nucleus by three dimensional imaging, whereas GFPs were not detected in any regions of cells. This micronucleus assay can readily be used in genetic based toxicity and efficacy assay to the various bio-chemicals and pharmaceutical evaluation.

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