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

Biochem Biophys Res Commun. 2016 Aug 5;476(4):580-585. doi: 10.1016/j.bbrc.2016.05.165. Epub 2016 May 31.

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.

Keywords: Diabetes; Encapsulation; Gene expression; Islet culture; Porcine islets.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Female
  • Gene Expression
  • Gene Ontology
  • Glucagon / metabolism
  • In Vitro Techniques
  • Insulin / metabolism
  • Insulin Secretion
  • Islets of Langerhans / metabolism*
  • Sepharose
  • Signal Transduction / genetics
  • Somatostatin / metabolism
  • Sus scrofa
  • Time Factors
  • Tissue Culture Techniques
  • Transcriptome

Substances

  • Insulin
  • Somatostatin
  • Glucagon
  • Sepharose