Live-imaging of Breast Epithelial Cell Migration After the Transient Depletion of TIP60

J Vis Exp. 2017 Dec 7:(130):56248. doi: 10.3791/56248.

Abstract

The wound-healing assay is efficient and one of the most economical ways to study cell migration in vitro. Conventionally, images are taken at the beginning and end of an experiment using a phase-contrast microscope, and the migration abilities of cells are evaluated by the closure of wounds. However, cell movement is a dynamic phenomenon, and a conventional method does not allow for tracking single-cell movement. To improve current wound-healing assays, we use live-cell imaging techniques to monitor cell migration in real time. This method allows us to determine the cell migration rate based on a cell tracking system and provides a clearer distinction between cell migration and cell proliferation. Here, we demonstrate the use of live-cell imaging in wound-healing assays to study the different migration abilities of breast epithelial cells influenced by the presence of TIP60. As cell motility is highly dynamic, our method provides more insights into the processes of wound healing than a snapshot of wound closure taken with the traditional imaging techniques used for wound-healing assays.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Breast / cytology*
  • Breast / diagnostic imaging*
  • Cell Movement / physiology*
  • Cell Proliferation / physiology
  • Epithelial Cells / cytology
  • Female
  • Humans
  • Lysine Acetyltransferase 5 / deficiency*
  • Lysine Acetyltransferase 5 / genetics
  • Lysine Acetyltransferase 5 / metabolism
  • Wound Healing

Substances

  • KAT5 protein, human
  • Lysine Acetyltransferase 5