Retention of gene expression in porcine islets after agarose encapsulation and long-term culture.
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.
The Rogosin Institute - Xenia Division, 740 Birch Road, Xenia, OH 45385, USA. Electronic address: pdumpala@rixd.org.
Pradeep R
PR
Dumpala
The Rogosin Institute - Xenia Division, 740 Birch Road, Xenia, OH 45385, USA.
Robert W
RW
Holdcraft
The Rogosin Institute - Xenia Division, 740 Birch Road, Xenia, OH 45385, USA.
Prithy C
PC
Martis
The Rogosin Institute - Xenia Division, 740 Birch Road, Xenia, OH 45385, USA.
Melissa A
MA
Laramore
The Rogosin Institute, 505 East 70th Street, New York, NY 10021, USA.
Thomas S
TS
Parker
The Rogosin Institute, 505 East 70th Street, New York, NY 10021, USA.
Daniel M
DM
Levine
The Rogosin Institute, 505 East 70th Street, New York, NY 10021, USA; NewYork-Presbyterian Hospital, Weill Medical College of Cornell University, 1300 York Avenue, New York, NY 10021, USA.
Barry H
BH
Smith
The Rogosin Institute - Xenia Division, 740 Birch Road, Xenia, OH 45385, USA.
Lawrence S
LS
Gazda
D000818:Animals; D005260:Female; D015870:Gene Expression; D063990:Gene Ontology; D005934:Glucagon / Q000378:metabolism; D066298:In Vitro Techniques; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000378:metabolism*; D012685:Sepharose; D015398:Signal Transduction / Q000235:genetics; D013004:Somatostatin / Q000378:metabolism; D034421:Sus scrofa; D013997:Time Factors; D046509:Tissue Culture Techniques; D059467:Transcriptome
D007328:Insulin; D013004:Somatostatin; D005934:Glucagon; D012685:Sepharose
Diabetes; Encapsulation; Gene expression; Islet culture; Porcine islets