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Title of Journal: Stem Cell Rev and Rep

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Abbravation: Stem Cell Reviews and Reports

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Effect of a Matrigel Sandwich on Endodermal Differentiation of Human Embryonic Stem Cells

Authors: Betty R. Lawton, Julie Ann Sosa, Sanziana Roman, Diane S. Krause,

Publish Date: 2013/05/30
Volume: 9, Issue:5, Pages: 578-585
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Definitive endoderm can be derived from human embryonic stem cells using low serum medium with cytokines involved in the epithelial-to-mesenchymal 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 endoderm-associated 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 endoderm.We 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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