Authors: Betty R Lawton Julie Ann Sosa Sanziana Roman Diane S Krause
Publish Date: 2013/05/30
Volume: 9, Issue: 5, Pages: 578-585
Abstract
Definitive endoderm can be derived from human embryonic stem cells using low serum medium with cytokines involved in the epithelialtomesenchymal transition including Activin A and Wnt3A The purpose of this study was to develop an improved protocol that permits the induction of definitive endoderm while avoiding the high rate of cell death that often occurs with existing protocols By including insulin and other nutrients we demonstrate that cell viability can be preserved throughout differentiation In addition modifying a matrigel sandwich method previously reported to induce precardiac mesoderm allows for enhanced endodermal differentiation based on expression of endodermassociated genes The morphological and migratory characteristics of cells cultured by the technique as well as gene expression patterns indicate that the protocol can emulate key events in gastrulation towards the induction of definitive endodermWe wish to acknowledge the generous contributions of Robert Epstein Peter Berman Laurence Weiss and the Burton Block Fund Funding was also received from Women’s Health Research at Yale and the Connecticut Stem Cell Research Fund We also wish to recognize the Yale Stem Cell Core the University of Connecticut Stem Cell Core and the Yale Stem Cell Center for advice and support
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