Construction of three quadruple-fluorescent MDA435 cell lines that enable monitoring of the whole chromosome segregation process in the living state

Mitotic events from prophase to telophase are defined by morphology or movement of chromatin, nuclear envelope, centrosomes and spindles. Live-cell imaging is useful for characterizing the whole chromosome segregation process in the living state. In this study, we constructed three quadruple-fluores...

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Published inMutation research Vol. 657; no. 1; pp. 56 - 62
Main Authors SUGIMOTO, Kenji, SENDA-MURATA, Kaori, OKA, Shigenori
Format Conference Proceeding Journal Article
LanguageEnglish
Published Amsterdam Elsevier 17.11.2008
Subjects
Biological and medical sciences
Biomarkers - analysis
Cell Line
Chromatin - ultrastructure
Chromosome Segregation
Fluorescence
Fundamental and applied biological sciences. Psychology
Genetics of eukaryotes. Biological and molecular evolution
Humans
Kinetochores - ultrastructure
Medical sciences
Mitosis
Models, Genetic
Nuclear Envelope - ultrastructure
Spindle Apparatus - ultrastructure
Toxicology
GFP
Live cell
Toxicology
Kinetochore
Cell line
Surveillance
Spindle
Segregation
Fluorescence
Chromosome
Genetics
Nuclear envelope
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Summary:Mitotic events from prophase to telophase are defined by morphology or movement of chromatin, nuclear envelope, centrosomes and spindles. Live-cell imaging is useful for characterizing the whole chromosome segregation process in the living state. In this study, we constructed three quadruple-fluorescent MDA435 cell lines in which chromatin, kinetochores, nuclear envelope and either inner centromere, microtubules or centrosomes/spindles were differentially visualized with cyan, green, orange and red fluorescent proteins (ECFP, EGFP, mKO and DsRed). Each mitotic stage of the individual cells could be identified by capturing live-cell images without the requirement of fixing or staining steps. In addition, we obtained four-color time-lapse images of one cell line, MDA-Auro/imp/H3/AF, from prophase to metaphase and from early anaphase to telophase. These quadruple-fluorescent cell lines will be useful for precisely analyzing the mitotic events from prophase through to telophase in single cells in the future.
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content type line 23
ISSN:1383-5718
0027-5107
1879-3592
DOI:10.1016/j.mrgentox.2008.08.005