Characterization of insulin-producing cells derived from PDX-1-transfected neural stem cells

Mol Med Rep. 2012 Dec;6(6):1428-32. doi: 10.3892/mmr.2012.1089. Epub 2012 Sep 20.

Abstract

Islet cell transplantation is a promising treatment strategy for type-1 diabetes. However, functional islet cells are hard to obtain for transplantation and are in short supply. Directing the differentiation of stem cells into insulin‑producing cells, which serve as islet cells, would overcome this shortage. Bone marrow contains hematopoietic stem cells and mesenchymal stem cells. The present study used bone marrow cells isolated from rats and neural stem cells (NSCs) that were derived from bone marrow cells in culture. Strong nestin staining was detected in NSCs, but not in bone marrow stromal cells (BMSCs). In vitro transfection of the pancreatic duodenal homeobox-1 (PDX-1) gene into NSCs generated insulin‑producing cells. Reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) analysis confirmed that PDX-1-transfected NSCs expressed insulin mRNA and released insulin protein. However, insulin release from PDX-1-transfected NSCs did not respond to the challenge of glucose and glucagon-like peptide-1. These results support the use of bone marrow-derived NSCs as a renewable source of insulin-producing cells for autologous transplantation to treat type-1 diabetes.

Keywords: bone marrow; nestin; neuron stem cell; pancreatic duodenal homeobox-1; insulin-producing cell; glucose.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bone Marrow Cells / cytology
  • Cell Differentiation
  • Cells, Cultured
  • Glucagon-Like Peptide 1 / pharmacology
  • Glucose / pharmacology
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism*
  • Insulin / genetics
  • Insulin / metabolism
  • Insulin-Secreting Cells / cytology*
  • Insulin-Secreting Cells / drug effects
  • Insulin-Secreting Cells / metabolism
  • Nestin / metabolism
  • Neural Stem Cells / cytology
  • Neural Stem Cells / metabolism*
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics
  • Trans-Activators / genetics
  • Trans-Activators / metabolism*
  • Transfection

Substances

  • Homeodomain Proteins
  • Insulin
  • Nestin
  • RNA, Messenger
  • Recombinant Fusion Proteins
  • Trans-Activators
  • enhanced green fluorescent protein
  • pancreatic and duodenal homeobox 1 protein
  • Green Fluorescent Proteins
  • Glucagon-Like Peptide 1
  • Glucose