期刊
ACTA BIOMATERIALIA
卷 22, 期 -, 页码 103-110出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.actbio.2015.04.013
关键词
Poly(ethylene glycol) hydrogels; Human embryonic stem cells; Pancreatic precursor cells; Long-term culture platforms; Controlled release
资金
- Beta Cell Biology Consortium
- National Institute of Diabetes and Digestive and Kidney Diseases [DK089561]
- U.S. Department of Education
- NSF CAREER [0847390]
- Direct For Mathematical & Physical Scien
- Division Of Materials Research [0847390] Funding Source: National Science Foundation
This study aimed to develop a three dimensional culture platform for aggregates of human embryonic stem cell (hESC)-derived pancreatic progenitors that enables long-term culture, maintains aggregate size and morphology, does not adversely affect differentiation and provides a means for aggregate recovery. A platform was developed with poly(ethylene glycol) hydrogels containing collagen type I, for cell-matrix interactions, and peptide crosslinkers, for facile recovery of aggregates. The platform was first demonstrated with RIN-m5F cells, showing encapsulation and subsequent release of single cells and aggregates without adversely affecting viability. Aggregates of hESC-derived pancreatic progenitors with an effective diameter of 82 (15) pm were either encapsulated in hydrogels or cultured in suspension for 28 days. At day 14, aggregate viability was maintained in the hydrogels, but significantly reduced (88%) in suspension culture. However by day 28, viability was reduced under both culture conditions. Aggregate size was maintained in the hydrogels, but in suspension was significantly higher (similar to 2-fold) by day 28. The ability to release aggregates followed by a second enzyme treatment to achieve single cells enabled assessment by flow cytometry. Prior to encapsulation, there were 39% Pdx1(+)/Nkx6.1(+) cells, key endocrine markers required for beta-cell maturation. The fraction of doubly positive cells was not affected in hydrogels but was slightly and significantly lower in suspension culture by 28 days. In conclusion, we demonstrate that a MMP-sensitive PEG hydrogel containing collagen type I is a promising platform for hESC-derived pancreatic progenitors that maintains viable aggregates, aggregate size, and progenitor state and offers facile recovery of aggregates. (C) 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
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