Journal
STEM CELL RESEARCH
Volume 6, Issue 3, Pages 276-285Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.scr.2011.02.004
Keywords
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Funding
- California Institute for Regenerative Medicine
- Juvenile Diabetes Research Foundation (JDRF) [3-2008-477, 35-2008-628, 1-2010-152]
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Pancreatic beta-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 beta-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 mu m 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 HNF1 beta-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. (C) 2011 Elsevier B.V. All rights reserved.
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