Unit

UNIT 14.9 Assessing Cell Fusion and Cytokinesis Failure as Mechanisms of Clone 9 Hepatocyte Multinucleation In Vitro

  1. Damir Simic,
  2. Catherine Euler,
  3. Christina Thurby,
  4. Mike Peden,
  5. Sarah Tannehill-Gregg,
  6. Todd Bunch,
  7. Thomas Sanderson,
  8. Terry Van Vleet

Published Online: 1 AUG 2012

DOI: 10.1002/0471140856.tx1409s53

Current Protocols in Toxicology

Current Protocols in Toxicology

How to Cite

Simic, D., Euler, C., Thurby, C., Peden, M., Tannehill-Gregg, S., Bunch, T., Sanderson, T. and Van Vleet, T. 2012. Assessing Cell Fusion and Cytokinesis Failure as Mechanisms of Clone 9 Hepatocyte Multinucleation In Vitro. Current Protocols in Toxicology. 53:14.9:14.9.1–14.9.17.

Author Information

  1. Drug Safety Evaluation, Bristol-Myers Squibb Co., Mount Vernon, Indiana

Publication History

  1. Published Online: 1 AUG 2012

Abstract

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. Curr. Protoc. Toxicol. 53:14.9.1-14.9.17. © 2012 by John Wiley & Sons, Inc.

Keywords:

  • multinucleation;
  • hepatocyte;
  • Clone 9;
  • cytokinesis;
  • cell fusion