Assessing cell fusion and cytokinesis failure as mechanisms of clone 9 hepatocyte multinucleation in vitro.
In this in vitro model of hepatocyte multinucleation, separate cultures of rat Clone 9 cells are labeled with either red or green cell tracker dyes (Red Cell Tracker CMPTX or Vybrant CFDA SE Cell Tracer), plated together in mixed-color colonies, and treated with positive or negative control agents for 4 days. The fluorescent dyes become cell-impermeant after entering cells and are not transferred to adjacent cells in a population, but are inherited by daughter cells after fusion. The mixed-color cultures are then evaluated microscopically for multinucleation and analysis of the underlying mechanism (cell fusion/cytokinesis). Multinucleated cells containing only one dye have undergone cytokinesis failure, whereas dual-labeled multinucleated cells have resulted from fusion.
Drug Safety Evaluation, Bristol-Myers Squibb Co, Mount Vernon, Indiana, USA.
Damir
D
Simic
Catherine
C
Euler
Christina
C
Thurby
Mike
M
Peden
Sarah
S
Tannehill-Gregg
Todd
T
Bunch
Thomas
T
Sanderson
Terry
T
Van Vleet
D000818:Animals; D001681:Biological Assay / Q000379:methods; D002459:Cell Fusion; D002478:Cells, Cultured; D002999:Clone Cells / Q000166:cytology* / Q000187:drug effects; D048749:Cytokinesis / Q000187:drug effects*; D005456:Fluorescent Dyes; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects*; D006822:Hybrid Cells / Q000187:drug effects*; D051381:Rats; D013194:Staining and Labeling / Q000379:methods*; D018675:Toxicity Tests / Q000379:methods; D015262:Xenobiotics / Q000633:toxicity
D005456:Fluorescent Dyes; D015262:Xenobiotics