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J Mol Endocrinol. 2016 Apr;56(3):201-11. doi: 10.1530/JME-15-0293. Epub 2016 Jan 27.

Peptide production and secretion in GLUTag, NCI-H716, and STC-1 cells: a comparison to native L-cells.

Author information

1
Department of Biomedical Sciences and NNF Center for Basic Metabolic Researchthe Panum Institute, University of Copenhagen, Copenhagen, Denmark.
2
Department of Clinical Biochemistry RigshospitaletUniversity of Copenhagen, Copenhagen, Denmark.
3
Cambridge Institute for Medical Research and MRC Metabolic Diseases UnitUniversity of Cambridge, Cambridge, United Kingdom.
4
Department of Biomedical Sciences and NNF Center for Basic Metabolic Researchthe Panum Institute, University of Copenhagen, Copenhagen, Denmark jjholst@sund.ku.dk.

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.

KEYWORDS:

GLP-1-producing cell lines; L-cells; peptide production

PMID:
26819328
DOI:
10.1530/JME-15-0293
[Indexed for MEDLINE]

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