title: Peptide production and secretion in GLUTag, NCI-H716, and STC-1 cells: a comparison to native L-cells.
abstract: GLUTag, NCI-H716, and STC-1 are cell lines that are widely used to study mechanisms underlying secretion of glucagon-like peptide-1 (GLP-1), but the extent to which they resemble native L-cells is unknown. We used validated immunoassays for 14 different hormones to analyze peptide content (lysis samples; n = 9 from different passage numbers) or peptide secretion in response to buffer (baseline), and after stimulation with 50 mM KCl or 10 mM glucose + 10 µM forskolin/3-isobutyl-1-methylxanthine (n = 6 also different passage numbers). All cell lines produced and processed proglucagon into GLP-1, GLP-2, glicentin, and oxyntomodulin in a pattern (prohormone convertase (PC)1/3 dependent) similar to that described for human gut. All three cell lines showed basal secretion of GLP-1 and GLP-2, which increased after stimulation. In contrast to freshly isolated murine L-cells, all cell lines also expressed PC2 and secreted large amounts of pancreatic glucagon. Neurotensin and somatostatin storage was low and secretion was not consistently increased by stimulation. STC-1 cells released more glucose-dependent insulinotropic polypeptide than GLP-1 at baseline (P < 0.01) and KCl elevated its secretion (P < 0.05). Peptide YY, which normally co-localizes with GLP-1 in distal L-cells, was not detected in any of the cell lines. GLUTag and STC-1 cells also expressed vasoactive intestinal peptide, but none expressed pancreatic polypeptide or insulin. GLUTag contained and secreted large amounts of CCK, while NCI-H716 did not store this peptide and STC-1 contained low amounts. Our results show that hormone production in cell line models of the L-cell has limited similarity to the natural L-cells.
authors.affiliation: Department of Biomedical Sciences and NNF Center for Basic Metabolic Researchthe Panum Institute, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Rune Ehrenreich
authors.initials: RE
authors.lastname: Kuhre
authors.affiliation: Department of Biomedical Sciences and NNF Center for Basic Metabolic Researchthe Panum Institute, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Nicolai Jacob
authors.initials: NJ
authors.lastname: Wewer Albrechtsen
authors.affiliation: Department of Biomedical Sciences and NNF Center for Basic Metabolic Researchthe Panum Institute, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Carolyn Fiona
authors.initials: CF
authors.lastname: Deacon
authors.affiliation: Department of Biomedical Sciences and NNF Center for Basic Metabolic Researchthe Panum Institute, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Emilie
authors.initials: E
authors.lastname: Balk-Møller
authors.affiliation: Department of Clinical Biochemistry RigshospitaletUniversity of Copenhagen, Copenhagen, Denmark.
authors.forename: Jens Frederik
authors.initials: JF
authors.lastname: Rehfeld
authors.affiliation: Cambridge Institute for Medical Research and MRC Metabolic Diseases UnitUniversity of Cambridge, Cambridge, United Kingdom.
authors.forename: Frank
authors.initials: F
authors.lastname: Reimann
authors.affiliation: Cambridge Institute for Medical Research and MRC Metabolic Diseases UnitUniversity of Cambridge, Cambridge, United Kingdom.
authors.forename: Fiona Mary
authors.initials: FM
authors.lastname: Gribble
authors.affiliation: Department of Biomedical Sciences and NNF Center for Basic Metabolic Researchthe Panum Institute, University of Copenhagen, Copenhagen, Denmark jjholst@sund.ku.dk.
authors.forename: Jens Juul
authors.initials: JJ
authors.lastname: Holst
mesh_terms: D000818:Animals; D002460:Cell Line; D052216:Glucagon-Like Peptide 1 / Q000378:metabolism*; D050416:Glucagon-Secreting Cells / Q000378:metabolism; D050417:Insulin-Secreting Cells / Q000378:metabolism; D051379:Mice; D010452:Peptide Biosynthesis; D010446:Peptide Fragments / Q000096:biosynthesis / Q000378:metabolism; D052336:Proglucagon / Q000096:biosynthesis / Q000737:chemistry
chemical_list: D010446:Peptide Fragments; D052336:Proglucagon; D052216:Glucagon-Like Peptide 1
keywords: GLP-1-producing cell lines; L-cells; peptide production
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