Document Detail

A microwell cell culture platform for the aggregation of pancreatic β-cells.
MedLine Citation:
PMID:  22320435     Owner:  NLM     Status:  MEDLINE    
Cell-cell contact between pancreatic β-cells is important for maintaining survival and normal insulin secretion. Various techniques have been developed to promote cell-cell contact between β-cells, but a simple yet robust method that affords precise control over three-dimensional (3D) β-cell cluster size has not been demonstrated. To address this need, we developed a poly(ethylene glycol) (PEG) hydrogel microwell platform using photolithography. This microwell cell-culture platform promotes the formation of 3D β-cell aggregates of defined sizes from 25 to 210 μm in diameter. Using this platform, mouse insulinoma 6 (MIN6) β-cells formed aggregates with cell-cell adherin junctions. These naturally formed cell aggregates with controllable sizes can be removed from the microwells for macroencapsulation, implantation, or other biological assays. When removed and subsequently encapsulated in PEG hydrogels, the aggregated cell clusters demonstrated improved cellular viability (>90%) over 7 days in culture, while the β-cells encapsulated as single cells maintained only 20% viability. Aggregated MIN6 cells also exhibited more than fourfold higher insulin secretion in response to a glucose challenge compared with encapsulated single β-cells. Further, the cell aggregates stained positively for E-cadherin, indicative of the formation of cell junctions. Using this hydrogel microwell cell-culture method, viable and functional β-cell aggregates of specific sizes were created, providing a platform from which other biologically relevant questions may be answered.
Abigail B Bernard; Chien-Chi Lin; Kristi S Anseth
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2012-03-19
Journal Detail:
Title:  Tissue engineering. Part C, Methods     Volume:  18     ISSN:  1937-3392     ISO Abbreviation:  Tissue Eng Part C Methods     Publication Date:  2012 Aug 
Date Detail:
Created Date:  2012-07-23     Completed Date:  2012-12-06     Revised Date:  2013-07-02    
Medline Journal Info:
Nlm Unique ID:  101466663     Medline TA:  Tissue Eng Part C Methods     Country:  United States    
Other Details:
Languages:  eng     Pagination:  583-92     Citation Subset:  IM    
Department of Chemical and Biological Engineering, University of Colorado, Boulder, CO 80309, USA.
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MeSH Terms
Cadherins / metabolism
Cell Communication
Cell Culture Techniques / methods*
Cell Survival
Cross-Linking Reagents / chemistry
Hydrogels / chemistry
Insulin / metabolism
Insulin-Secreting Cells / cytology*
Microscopy, Confocal / methods
Microscopy, Fluorescence / methods
Polyethylene Glycols / chemistry
Polymers / chemistry
Grant Support
R01DK076084/DK/NIDDK NIH HHS; //Howard Hughes Medical Institute
Reg. No./Substance:
0/Cadherins; 0/Cross-Linking Reagents; 0/Hydrogels; 0/Insulin; 0/Polyethylene Glycols; 0/Polymers

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