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Title: Retention of gene expression in porcine islets after agarose encapsulation and long-term culture

Abstract

Agarose encapsulation of porcine islets allows extended in vitro culture, providing ample time to determine the functional capacity of the islets and conduct comprehensive microbiological safety testing prior to implantation as a treatment for type 1 diabetes mellitus. However, the effect that agarose encapsulation and long-term culture may have on porcine islet gene expression is unknown. The aim of the present study was to compare the transcriptome of encapsulated porcine islets following long-term in vitro culture against free islets cultured overnight. Global gene expression analysis revealed no significant change in the expression of 98.47% of genes. This indicates that the gene expression profile of free islets is highly conserved following encapsulation and long-term culture. Importantly, the expression levels of genes that code for critical hormones secreted by islets (insulin, glucagon, and somatostatin) as well as transcripts encoding proteins involved in their packaging and secretion are unchanged. While a small number of genes known to play roles in the insulin secretion and insulin signaling pathways are differentially expressed, our results show that overall gene expression is retained following islet isolation, agarose encapsulation, and long-term culture. - Highlights: • Effect of agarose encapsulation and 8 week culture on porcine islets was analyzed. • Transcriptomemore » analysis revealed no significant change in a majority (98%) of genes. • Agarose encapsulation allows for long-term culture of porcine islets. • Islet culture allows for functional and microbial testing prior to clinical use.« less

Authors:
 [1]; ; ;  [1]; ;  [2];  [2];  [3];  [1]
  1. The Rogosin Institute – Xenia Division, 740 Birch Road, Xenia, OH 45385 (United States)
  2. The Rogosin Institute, 505 East 70th Street, New York, NY 10021 (United States)
  3. (United States)
Publication Date:
OSTI Identifier:
22606139
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 476; Journal Issue: 4; Other Information: Copyright (c) 2016 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; DIABETES MELLITUS; ENCAPSULATION; GENES; GLUCAGON; IN VITRO; INSULIN; RETENTION; SAFETY; SECRETION; SOMATOSTATIN

Citation Formats

Dumpala, Pradeep R., E-mail: pdumpala@rixd.org, Holdcraft, Robert W., Martis, Prithy C., Laramore, Melissa A., Parker, Thomas S., Levine, Daniel M., Smith, Barry H., NewYork-Presbyterian Hospital, Weill Medical College of Cornell University, 1300 York Avenue, New York, NY 10021, and Gazda, Lawrence S. Retention of gene expression in porcine islets after agarose encapsulation and long-term culture. United States: N. p., 2016. Web. doi:10.1016/J.BBRC.2016.05.165.
Dumpala, Pradeep R., E-mail: pdumpala@rixd.org, Holdcraft, Robert W., Martis, Prithy C., Laramore, Melissa A., Parker, Thomas S., Levine, Daniel M., Smith, Barry H., NewYork-Presbyterian Hospital, Weill Medical College of Cornell University, 1300 York Avenue, New York, NY 10021, & Gazda, Lawrence S. Retention of gene expression in porcine islets after agarose encapsulation and long-term culture. United States. doi:10.1016/J.BBRC.2016.05.165.
Dumpala, Pradeep R., E-mail: pdumpala@rixd.org, Holdcraft, Robert W., Martis, Prithy C., Laramore, Melissa A., Parker, Thomas S., Levine, Daniel M., Smith, Barry H., NewYork-Presbyterian Hospital, Weill Medical College of Cornell University, 1300 York Avenue, New York, NY 10021, and Gazda, Lawrence S. Fri . "Retention of gene expression in porcine islets after agarose encapsulation and long-term culture". United States. doi:10.1016/J.BBRC.2016.05.165.
@article{osti_22606139,
title = {Retention of gene expression in porcine islets after agarose encapsulation and long-term culture},
author = {Dumpala, Pradeep R., E-mail: pdumpala@rixd.org and Holdcraft, Robert W. and Martis, Prithy C. and Laramore, Melissa A. and Parker, Thomas S. and Levine, Daniel M. and Smith, Barry H. and NewYork-Presbyterian Hospital, Weill Medical College of Cornell University, 1300 York Avenue, New York, NY 10021 and Gazda, Lawrence S.},
abstractNote = {Agarose encapsulation of porcine islets allows extended in vitro culture, providing ample time to determine the functional capacity of the islets and conduct comprehensive microbiological safety testing prior to implantation as a treatment for type 1 diabetes mellitus. However, the effect that agarose encapsulation and long-term culture may have on porcine islet gene expression is unknown. The aim of the present study was to compare the transcriptome of encapsulated porcine islets following long-term in vitro culture against free islets cultured overnight. Global gene expression analysis revealed no significant change in the expression of 98.47% of genes. This indicates that the gene expression profile of free islets is highly conserved following encapsulation and long-term culture. Importantly, the expression levels of genes that code for critical hormones secreted by islets (insulin, glucagon, and somatostatin) as well as transcripts encoding proteins involved in their packaging and secretion are unchanged. While a small number of genes known to play roles in the insulin secretion and insulin signaling pathways are differentially expressed, our results show that overall gene expression is retained following islet isolation, agarose encapsulation, and long-term culture. - Highlights: • Effect of agarose encapsulation and 8 week culture on porcine islets was analyzed. • Transcriptome analysis revealed no significant change in a majority (98%) of genes. • Agarose encapsulation allows for long-term culture of porcine islets. • Islet culture allows for functional and microbial testing prior to clinical use.},
doi = {10.1016/J.BBRC.2016.05.165},
journal = {Biochemical and Biophysical Research Communications},
number = 4,
volume = 476,
place = {United States},
year = {Fri Aug 05 00:00:00 EDT 2016},
month = {Fri Aug 05 00:00:00 EDT 2016}
}