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Abstract

Biology

Measuring the Confluence of iPSCs Using an Automated Imaging System

Published: June 10th, 2020

DOI:

10.3791/61225

1Genetics and Rare Diseases Research Division, Ospedale Pediatrico Bambino Gesù, IRCCS, 2Department of Science, University Roma Tre, 3Department of Onco-hematology, Gene and Cell Therapy, Ospedale Pediatrico Bambino Gesù, IRCCS

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.

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Keywords IPSCs

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