Measuring the Confluence of iPSCs using an Automated Imaging System

J Vis Exp. 2020 Jun 10:(160). doi: 10.3791/61225.

Abstract

This study focuses on understanding how growing iPSCs on different ECM coating substrates can affect cell confluence. A protocol to assess iPSC confluence in real time has been established without the need to count cells in single cell suspension to avoid any growth perturbation. A high-content image analysis system was used to assess iPCS confluence on 4 different ECMs over time in an automated manner. Different analysis settings were used to assess cell confluence of adherent iPSCs and only a slight difference (at 24 and 48 hours with laminin) has been observed whether a 60, 80 or 100% mask was applied. We also show that laminin lead to the best confluence compared to Matrigel, vitronectin and fibronectin.

Publication types

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

MeSH terms

  • Automation
  • Cell Count
  • Cytoskeleton / metabolism
  • Extracellular Matrix / metabolism
  • Humans
  • Image Processing, Computer-Assisted*
  • Induced Pluripotent Stem Cells / cytology*