Isolation of Mouse Pancreatic Alpha, Beta, Duct and Acinar Populations with Cell Surface Markers

Overview

Journal Mol Cell Endocrinol

Specialties Cell Biology
Endocrinology
Molecular Biology

Date 2011 May 5

PMID 21539888

Citations 30

Authors

Craig Dorrell

Maria T Grompe

Fong Cheng Pan

Yongping Zhong

Pamela S Canaday

Leonard D Shultz

Dale L Greiner

Chris V Wright

Philip R Streeter

Markus Grompe

Affiliations

Soon will be listed here.

Abstract

Tools permitting the isolation of live pancreatic cell subsets for culture and/or molecular analysis are limited. To address this, we developed a collection of monoclonal antibodies with selective surface labeling of endocrine and exocrine pancreatic cell types. Cell type labeling specificity and cell surface reactivity were validated on mouse pancreatic sections and by gene expression analysis of cells isolated using FACS. Five antibodies which marked populations of particular interest were used to isolate and study viable populations of purified pancreatic ducts, acinar cells, and subsets of acinar cells from whole pancreatic tissue or of alpha or beta cells from isolated mouse islets. Gene expression analysis showed the presence of known endocrine markers in alpha and beta cell populations and revealed that TTR and DPPIV are primarily expressed in alpha cells whereas DGKB and GPM6A have a beta cell specific expression profile.

Citing Articles

Identification of a Novel Subset of Human Airway Epithelial Basal Stem Cells.

Cheng C, Katoch P, Zhong Y, Higgins C, Moredock M, Chang M Int J Mol Sci. 2024; 25(18).

PMID: 39337350 PMC: 11432080. DOI: 10.3390/ijms25189863.

3D chromatin maps of the human pancreas reveal lineage-specific regulatory architecture of T2D risk.

Su C, Gao L, May C, Pippin J, Boehm K, Lee M Cell Metab. 2022; 34(9):1394-1409.e4.

PMID: 36070683 PMC: 9664375. DOI: 10.1016/j.cmet.2022.08.014.

Variant-to-gene-mapping analyses reveal a role for pancreatic islet cells in conferring genetic susceptibility to sleep-related traits.

Lasconi C, Pahl M, Pippin J, Su C, Johnson M, Chesi A Sleep. 2022; 45(8).

PMID: 35537191 PMC: 9366645. DOI: 10.1093/sleep/zsac109.

Protein Kinase C (Pkc)-δ Mediates Arginine-Induced Glucagon Secretion in Pancreatic α-Cells.

Honzawa N, Fujimoto K, Kobayashi M, Kohno D, Kikuchi O, Yokota-Hashimoto H Int J Mol Sci. 2022; 23(7).

PMID: 35409362 PMC: 8999522. DOI: 10.3390/ijms23074003.

Pleiotrophin Expression and Actions in Pancreatic β-Cells.

Sevillano J, Liang A, Strutt B, Hill T, Szlapinski S, Ramos-Alvarez M Front Endocrinol (Lausanne). 2022; 13:777868.

PMID: 35250852 PMC: 8894601. DOI: 10.3389/fendo.2022.777868.

References

Maric D, Barker J . Neural stem cells redefined: a FACS perspective. Mol Neurobiol. 2004; 30(1):49-76. DOI: 10.1385/MN:30:1:049. View

Swart B, Salganik M, Wand M, Tinckam K, Milford E, Drbal K . The HLDA8 blind panel: findings and conclusions. J Immunol Methods. 2005; 305(1):75-83. DOI: 10.1016/j.jim.2005.07.003. View

Xia B, Zhan X, Yi R, Yang B . Can pancreatic duct-derived progenitors be a source of islet regeneration?. Biochem Biophys Res Commun. 2009; 383(4):383-5. DOI: 10.1016/j.bbrc.2009.03.114. View

Quoix N, Cheng-Xue R, Guiot Y, Herrera P, Henquin J, Gilon P . The GluCre-ROSA26EYFP mouse: a new model for easy identification of living pancreatic alpha-cells. FEBS Lett. 2007; 581(22):4235-40. DOI: 10.1016/j.febslet.2007.07.068. View

Dorrell C, Erker L, Lanxon-Cookson K, Abraham S, Victoroff T, Ro S . Surface markers for the murine oval cell response. Hepatology. 2008; 48(4):1282-91. PMC: 3121577. DOI: 10.1002/hep.22468. View

Basma H, Soto-Gutierrez A, Yannam G, Liu L, Ito R, Yamamoto T . Differentiation and transplantation of human embryonic stem cell-derived hepatocytes. Gastroenterology. 2008; 136(3):990-9. PMC: 2732349. DOI: 10.1053/j.gastro.2008.10.047. View

Gadue P, Gouon-Evans V, Cheng X, Wandzioch E, Zaret K, Grompe M . Generation of monoclonal antibodies specific for cell surface molecules expressed on early mouse endoderm. Stem Cells. 2009; 27(9):2103-13. PMC: 2890285. DOI: 10.1002/stem.147. View

Zhou Q, Brown J, Kanarek A, Rajagopal J, Melton D . In vivo reprogramming of adult pancreatic exocrine cells to beta-cells. Nature. 2008; 455(7213):627-32. PMC: 9011918. DOI: 10.1038/nature07314. View

Kikugawa R, Katsuta H, Akashi T, Yatoh S, Weir G, Sharma A . Differentiation of COPAS-sorted non-endocrine pancreatic cells into insulin-positive cells in the mouse. Diabetologia. 2009; 52(4):645-52. PMC: 4336153. DOI: 10.1007/s00125-009-1260-8. View

10.

Narushima M, Kobayashi N, Okitsu T, Tanaka Y, Li S, Chen Y . A human beta-cell line for transplantation therapy to control type 1 diabetes. Nat Biotechnol. 2005; 23(10):1274-82. DOI: 10.1038/nbt1145. View

11.

Lukowiak B, Vandewalle B, Riachy R, Kerr-Conte J, Gmyr V, Belaich S . Identification and purification of functional human beta-cells by a new specific zinc-fluorescent probe. J Histochem Cytochem. 2001; 49(4):519-28. DOI: 10.1177/002215540104900412. View

12.

Dorrell C, Abraham S, Lanxon-Cookson K, Canaday P, Streeter P, Grompe M . Isolation of major pancreatic cell types and long-term culture-initiating cells using novel human surface markers. Stem Cell Res. 2009; 1(3):183-94. DOI: 10.1016/j.scr.2008.04.001. View

13.

Michibata H, Okuno T, Konishi N, Wakimoto K, Kyono K, Aoki K . Inhibition of mouse GPM6A expression leads to decreased differentiation of neurons derived from mouse embryonic stem cells. Stem Cells Dev. 2008; 17(4):641-51. DOI: 10.1089/scd.2008.0088. View

14.

Westermark G, Westermark P . Transthyretin and amyloid in the islets of Langerhans in type-2 diabetes. Exp Diabetes Res. 2008; 2008:429274. PMC: 2553203. DOI: 10.1155/2008/429274. View

15.

Lawson D, Xin L, Lukacs R, Cheng D, Witte O . Isolation and functional characterization of murine prostate stem cells. Proc Natl Acad Sci U S A. 2006; 104(1):181-6. PMC: 1716155. DOI: 10.1073/pnas.0609684104. View

16.

Dupuis J, Langenberg C, Prokopenko I, Saxena R, Soranzo N, Jackson A . New genetic loci implicated in fasting glucose homeostasis and their impact on type 2 diabetes risk. Nat Genet. 2010; 42(2):105-16. PMC: 3018764. DOI: 10.1038/ng.520. View

17.

Hara M, Wang X, Kawamura T, Bindokas V, Dizon R, Alcoser S . Transgenic mice with green fluorescent protein-labeled pancreatic beta -cells. Am J Physiol Endocrinol Metab. 2002; 284(1):E177-83. DOI: 10.1152/ajpendo.00321.2002. View

18.

Williams C, Stechmann C, McLoon S . Subtractive immunization techniques for the production of monoclonal antibodies to rare antigens. Biotechniques. 1992; 12(6):842-7. View

19.

Flatt P, Bailey C, Green B . Dipeptidyl peptidase IV (DPP IV) and related molecules in type 2 diabetes. Front Biosci. 2008; 13:3648-60. DOI: 10.2741/2956. View

20.

Noguchi H, Xu G, Matsumoto S, Kaneto H, Kobayashi N, Bonner-Weir S . Induction of pancreatic stem/progenitor cells into insulin-producing cells by adenoviral-mediated gene transfer technology. Cell Transplant. 2007; 15(10):929-38. DOI: 10.3727/000000006783981431. View