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Stem Cell Res. 2011 May;6(3):276-85. doi: 10.1016/j.scr.2011.02.004. Epub 2011 Mar 2.

Differentiation of human embryonic stem cells into pancreatic endoderm in patterned size-controlled clusters.

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  • 1Diabetes Center, University of California San Francisco, 35 Medical Center Way, San Francisco, CA 94143, USA.

Abstract

Pancreatic β-cells function optimally when clustered in islet-like structures. However, nutrient and oxygen deprivation limits the viability of cells at the core of excessively large clusters. Hence, production of functional β-cells from human embryonic stem cells (hESCs) for patients with diabetes would benefit from the growth and differentiation of these cells in size-controlled aggregates. In this study, we controlled cluster size by seeding hESCs onto glass cover slips patterned by the covalent microcontact-printing of laminin in circular patches of 120 μm in diameter. These were used as substrates to grow and differentiate hESCs first into SOX17-positive/SOX7-negative definitive endoderm, after which many clusters released and formed uniformly sized three-dimensional clusters. Both released clusters and those that remained attached differentiated into HNF1β-positive primitive gut tube-like cells with high efficiency. Further differentiation yielded pancreatic endoderm-like cells that co-expressed PDX1 and NKX6.1. Controlling aggregate size allows efficient production of uniformly-clustered pancreatic endocrine precursors for in vivo engraftment or further in vitro maturation.

Copyright © 2011 Elsevier B.V. All rights reserved.

PMID:
21513906
[PubMed - indexed for MEDLINE]
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