Glucagon-like peptide-1 enhances production of insulin in insulin-producing cells derived from mouse embryonic stem cells

• Published in: Journal of endocrinology Vol. 186; no. 2; pp. 343 - 352
• Main Authors: Bai, L, Meredith, G, Tuch, B E
• Format: Journal Article
• Language: English
• Published: Colchester BioScientifica 01.08.2005; Portland Press
• Subjects: Animals; Biological and medical sciences; C-Peptide - analysis; C-Peptide - genetics; C-Peptide - metabolism; Cell Differentiation; Cell Line; Endocrine pancreas; Exenatide; Fundamental and applied biological sciences. Psychology; Glucagon - pharmacology; Glucagon-Like Peptide 1; Glucagon-Like Peptide-1 Receptor; Hormones. Régulation; Immunohistochemistry - methods; Insulin - biosynthesis; Insulin - genetics; Insulin - metabolism; Insulin Secretion; Mice; Peptide Fragments - pharmacology; Peptides - pharmacology; Protein Precursors - pharmacology; Receptors, Glucagon - metabolism; Regular papers; Reverse Transcriptase Polymerase Chain Reaction; Stem Cells - drug effects; Stem Cells - metabolism; Venoms - pharmacology; Vertebrates: endocrinology; Pancreatic hormone; Vertebrata; Gastrointestinal hormone; Mammalia; Mouse; Stem cell; Rodentia; Biosynthesis; Differentiation; In vitro; Insulin; Glucagon like peptide 1
• ISSN: 0022-0795; 1479-6805
• DOI: 10.1677/joe.1.06078

Embryonic stem cells (ESCs) can be differentiated into insulin-producing cells by a five-stage procedure involving altering culture conditions and addition of nicotinamide. The amounts of insulin in these cells are lower than those found in pancreatic β cells. Glucagon-like peptide-1 (GLP-1) induces the differentiation of β cells from ductal progenitor cells. We examined the possibility of GLP-1, and its long-acting agonist exendin-4, enhancing the differentiation of insulin-producing cells from mouse ESCs (mESCs). A five-stage culturing strategy starting with embryoid bodies (EBs) was used in this study. mRNA for pancreatic duodenal homeobox gene 1 (PDX-1) and neurogenic differentiation (NeuroD) was detected from stage 1, hepatocyte nuclear factor 3 beta (HNF3β) and insulin 2 from stage 2, Ngn3 and glucose transporter 2 (GLUT2) from stage 3, and insulin 1 and other β-cell markers, at stages 4–5. Cells at stage 5 secreted C-peptide, being 0.68 ± 0.01 pmol/106 cells per 2 days, and had an immunoreactive insulin content of 13.5 ± 0.7 pmol/106 cells. Addition of GLP-1 (100 nM) and nicotinamide (10 mM) at stage 5 resulted in a 50% and 48% increase in insulin content and C-peptide secretion respectively compared with nicotinamide alone. Glucose-induced insulin secretion was enhanced 4-fold by addition of both growth factors. The GLP-1 receptor was present at all five stages of the culture. Addition of exendin-4 to cells at stage 2 resulted in a 4.9-fold increase in expression of the gene for insulin 1 and a 2-fold increase in insulin content compared with the effect of nicotinamide alone at stage 5. It is concluded that both GLP-1 and exendin-4 enhance the level of expression of insulin in glucose-responsive insulin-producing cells derived from the R1 mESC line.