Live Cell Imaging with Time Lapse Photography to Study Epidermal Keratinocyte Proliferation Kinetics

Live-cell imaging is an evolving and somewhat challenging method to study keratinocyte behavior in vitro. Historically, keratinocyte division behavior was investigated via methods such as clonal analysis, immunostaining, and cell cycle analysis. None of these methods allow for the analysis of keratinocyte behavior at the single-cell level in real time. Over the past decade, groups have utilized live cell imaging to identify keratinocyte stem cells and committed progenitors without the need for labeling. Differences have been identified in each respective group's division behavior, rate of terminal differentiation, and cell cycle duration. Here, a method for keratinocyte live cell imaging with time-lapse photography and its analysis is described. Utilizing unpassaged keratinocytes is recommended for this method to most closely mimic in vivo behavior. Live cell imaging provides a unique ability to study stem cell and committed progenitor behavior at the single cell level and to determine division fates, cell cycle duration, as well as other proliferation metrics.