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.
Clone Cells/cytology , Cytokinesis/drug effects , Fluorescent Dyes , Hepatocytes/drug effects , Hybrid Cells/drug effects , Staining and Labeling/methods , Animals , Biological Assay/methods , Cell Fusion , Cells, Cultured , Clone Cells/drug effects , Hepatocytes/cytology , Rats , Toxicity Tests/methods , Xenobiotics/toxicity
