Proliferation of Sorted Human and Rat Beta Cells

Overview

Journal Diabetologia

Specialty Endocrinology

Date 2007 Nov 13

PMID 17994216

Citations 125

Authors

G Parnaud

D Bosco

T Berney

F Pattou

J Kerr-Conte

M Y Donath

C Bruun

T Mandrup-Poulsen

N Billestrup

P A Halban

Affiliations

Soon will be listed here.

Abstract

Aims/hypothesis: The aim of the study was to determine whether purified beta cells can replicate in vitro and whether this is enhanced by extracellular matrix (ECM) and growth factors.

Methods: Human beta cells were purified by FACS by virtue of their high zinc content using Newport Green, and excluding ductal and dead cells. Rat beta cells were sorted by autofluorescence or using the same method developed for human cells. Cells were plated on poly-L-lysine or ECMs from rat or human bladder carcinoma cells or bovine corneal ECM and incubated in the presence of BrdU with or without growth factors.

Results: The newly developed method for sorting human beta cells yields a population containing 91.4 +/- 2.8% insulin-positive cells with a low level of spontaneous apoptosis and a robust secretory response to glucose. Beta cells from 8-week-old rats proliferated in culture and this was increased by ECM. Among growth factors, only human growth hormone (hGH) and the glucagon-like peptide-1 analogue liraglutide enhanced proliferation of rat beta cells, with a significant increase on both poly-L-lysine and ECM. By contrast, sorted adult human beta cells from 16 donors aged 48.9 +/- 14.3 years (range 16-64 years) failed to replicate demonstrably in vitro regardless of the substratum or growth factors used.

Conclusions/interpretation: These findings indicate that, in our conditions, the fully differentiated human adult insulin-producing beta cell was unable to proliferate in vitro. This has important implications for any attempt to expand cells from pancreases of donors of this age group. By contrast, the rat beta cells used here were able to divide in vitro, and this was enhanced by ECM, hGH and liraglutide.

Citing Articles

DECORIN, a triceps-derived myokine, protects sorted β-cells and human islets against chronic inflammation associated with type 2 diabetes.

Langlois A, Cherfan J, Meugnier E, Rida A, Arous C, Peronet C Acta Physiol (Oxf). 2025; 241(2):e14267.

PMID: 39844653 PMC: 11754997. DOI: 10.1111/apha.14267.

Normoglycemia and physiological cortisone level maintain glucose homeostasis in a pancreas-liver microphysiological system.

Rigal S, Casas B, Kanebratt K, Wennberg Huldt C, Magnusson L, Mullers E Commun Biol. 2024; 7(1):877.

PMID: 39025915 PMC: 11258270. DOI: 10.1038/s42003-024-06514-w.

Advanced Imaging Techniques for the Characterization of Subcellular Organelle Structure in Pancreatic Islet β Cells.

McLaughlin M, Weaver S, Syed F, Evans-Molina C Compr Physiol. 2023; 14(1):5243-5267.

PMID: 38158370 PMC: 11490899. DOI: 10.1002/cphy.c230002.

FALCON systematically interrogates free fatty acid biology and identifies a novel mediator of lipotoxicity.

Wieder N, Fried J, Kim C, Sidhom E, Brown M, Marshall J Cell Metab. 2023; 35(5):887-905.e11.

PMID: 37075753 PMC: 10257950. DOI: 10.1016/j.cmet.2023.03.018.

FALCON systematically interrogates free fatty acid biology and identifies a novel mediator of lipotoxicity.

Wieder N, Fried J, Kim C, Sidhom E, Brown M, Marshall J bioRxiv. 2023; .

PMID: 36865221 PMC: 9979987. DOI: 10.1101/2023.02.19.529127.

References

Kieffer T, Habener J . The glucagon-like peptides. Endocr Rev. 1999; 20(6):876-913. DOI: 10.1210/edrv.20.6.0385. View

Chase L, Ulloa-Montoya F, Kidder B, Verfaillie C . Islet-derived fibroblast-like cells are not derived via epithelial-mesenchymal transition from Pdx-1 or insulin-positive cells. Diabetes. 2006; 56(1):3-7. DOI: 10.2337/db06-1165. View

Demeterco C, Beattie G, Dib S, Lopez A, Hayek A . A role for activin A and betacellulin in human fetal pancreatic cell differentiation and growth. J Clin Endocrinol Metab. 2000; 85(10):3892-7. DOI: 10.1210/jcem.85.10.6848. View

Beattie G, Montgomery A, Lopez A, Hao E, Perez B, Just M . A novel approach to increase human islet cell mass while preserving beta-cell function. Diabetes. 2002; 51(12):3435-9. DOI: 10.2337/diabetes.51.12.3435. View

Sutton R, Peters M, McShane P, Gray D, Morris P . Isolation of rat pancreatic islets by ductal injection of collagenase. Transplantation. 1986; 42(6):689-91. DOI: 10.1097/00007890-198612000-00022. View

Lefebvre V, Otonkoski T, Ustinov J, Huotari M, Pipeleers D, Bouwens L . Culture of adult human islet preparations with hepatocyte growth factor and 804G matrix is mitogenic for duct cells but not for beta-cells. Diabetes. 1998; 47(1):134-7. DOI: 10.2337/diab.47.1.134. View

Weinberg N, Ouziel-Yahalom L, Knoller S, Efrat S, Dor Y . Lineage tracing evidence for in vitro dedifferentiation but rare proliferation of mouse pancreatic beta-cells. Diabetes. 2007; 56(5):1299-304. DOI: 10.2337/db06-1654. View

Adams J, Watt F . Regulation of development and differentiation by the extracellular matrix. Development. 1993; 117(4):1183-98. DOI: 10.1242/dev.117.4.1183. View

Gmyr V, Kerr-Conte J, Belaich S, Vandewalle B, Leteurtre E, Vantyghem M . Adult human cytokeratin 19-positive cells reexpress insulin promoter factor 1 in vitro: further evidence for pluripotent pancreatic stem cells in humans. Diabetes. 2000; 49(10):1671-80. DOI: 10.2337/diabetes.49.10.1671. View

10.

Teta M, Long S, Wartschow L, Rankin M, Kushner J . Very slow turnover of beta-cells in aged adult mice. Diabetes. 2005; 54(9):2557-67. DOI: 10.2337/diabetes.54.9.2557. View

11.

Lelievre S, Weaver V, Bissell M . Extracellular matrix signaling from the cellular membrane skeleton to the nuclear skeleton: a model of gene regulation. Recent Prog Horm Res. 1996; 51:417-32. PMC: 2937008. View

12.

Van De Winkel M, Smets G, GEPTS W, Pipeleers D . Islet cell surface antibodies from insulin-dependent diabetics bind specifically to pancreatic B cells. J Clin Invest. 1982; 70(1):41-9. PMC: 370224. DOI: 10.1172/jci110601. View

13.

Billestrup N, Nielsen J . The stimulatory effect of growth hormone, prolactin, and placental lactogen on beta-cell proliferation is not mediated by insulin-like growth factor-I. Endocrinology. 1991; 129(2):883-8. DOI: 10.1210/endo-129-2-883. View

14.

Beattie G, Itkin-Ansari P, Cirulli V, Leibowitz G, Lopez A, Bossie S . Sustained proliferation of PDX-1+ cells derived from human islets. Diabetes. 1999; 48(5):1013-9. DOI: 10.2337/diabetes.48.5.1013. View

15.

Roskelley C, Srebrow A, Bissell M . A hierarchy of ECM-mediated signalling regulates tissue-specific gene expression. Curr Opin Cell Biol. 1995; 7(5):736-47. PMC: 2933201. DOI: 10.1016/0955-0674(95)80117-0. View

16.

Li S, Edgar D, Fassler R, Wadsworth W, Yurchenco P . The role of laminin in embryonic cell polarization and tissue organization. Dev Cell. 2003; 4(5):613-24. DOI: 10.1016/s1534-5807(03)00128-x. View

17.

Parnaud G, Hammar E, Rouiller D, Armanet M, Halban P, Bosco D . Blockade of beta1 integrin-laminin-5 interaction affects spreading and insulin secretion of rat beta-cells attached on extracellular matrix. Diabetes. 2006; 55(5):1413-20. DOI: 10.2337/db05-1388. View

18.

Drucker D . Glucagon-like peptides: regulators of cell proliferation, differentiation, and apoptosis. Mol Endocrinol. 2003; 17(2):161-71. DOI: 10.1210/me.2002-0306. View

19.

Sorenson R, Brelje T . Adaptation of islets of Langerhans to pregnancy: beta-cell growth, enhanced insulin secretion and the role of lactogenic hormones. Horm Metab Res. 1997; 29(6):301-7. DOI: 10.1055/s-2007-979040. View

20.

Rouiller D, Cirulli V, Halban P . Differences in aggregation properties and levels of the neural cell adhesion molecule (NCAM) between islet cell types. Exp Cell Res. 1990; 191(2):305-12. DOI: 10.1016/0014-4827(90)90019-7. View