Three Dimensional Nucleus Imaging analysis &amp; in vitro Genotoxicity Test through Advanced Cytokinesis-Block Micronucleus Assay (진보된 세포분열억제 소핵 평가법을 활용한 체외 유전독성검사 및 3 차원 핵 이미지 분석)

Hye-Ji Kim (김혜지); In-Cheol Seo (서인철); Whui-Su Shim (심휘수); Chae-Eun Lee (이채은); Won-Seok Chang (장원석); Hyun-Su Lee (이현수); Sun-Hyun Park (박순현)

doi:10.23032/jaae.2018.12.1.001

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.

KEYWORDS

The cytokinesis-block micronucleus cytome assay, micronuclei (MNi), advanced cytokinesis-block micronucleus (ACBMN) assay DAPI staining, TANNylated GFPs, Paraformaldehyde (PFA), Three dimensional image

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Journal of Alternatives to Animal Experiments 2023 KCI Impact Factor : 0.07

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

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* References for papers published after 2023 are currently being built.

Journal of Alternatives to Animal Experiments 2023 KCI Impact Factor : 0.07

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

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