Differentiation of COPAS-sorted Non-endocrine Pancreatic Cells into Insulin-positive Cells in the Mouse

Overview

Journal Diabetologia

Specialty Endocrinology

Date 2009 Feb 3

PMID 19183938

Citations 9

Authors

R Kikugawa

H Katsuta

T Akashi

S Yatoh

G C Weir

A Sharma

S Bonner-Weir

Affiliations

Soon will be listed here.

Abstract

Aims/hypothesis: The regenerative process in the pancreas is of particular interest, since insulin-producing beta cells are lost in diabetes. Differentiation of new beta cells from pancreatic non-endocrine cells has been reported in vivo and in vitro, a finding that implies the existence of pancreatic stem/progenitor cells. However, while tissue-specific stem cells are well documented in skin, intestine and testis, pancreatic stem cells have been elusive. We hypothesised that pancreatic stem/progenitor cells within the non-endocrine fraction could be a source of new islets in vitro.

Methods: To test if there were such cells within the pancreas, we generated pancreatic cell aggregates from tissue remaining after islet isolation from mouse insulin promoter 1-green fluorescent protein (MIP-GFP) mice. To eliminate any contamination of insulin-positive cells, we deleted all GFP-positive aggregates using COPAS Select and cultured with Matrigel. Immunohistochemistry, quantitative real-time PCR and single-cell nested RT-PCR were performed to confirm formation of insulin-producing cells.

Results: The GFP-negative cells were expanded as monolayers and then differentiated into three-dimensional cystic structures. After 1 week of culture, GFP-positive cells were found as clusters or single cells. By quantitative real-time PCR, no insulin mRNA was detected immediately after COPAS sorting, but after differentiation insulin mRNA of the whole preparation was 1.91 +/- 0.31% that of purified MIP-GFP beta cells. All GFP-positive cells expressed insulin 1; most expressed insulin 2, pancreas duodenum homeobox-1 and cytokeratin 19 by single cell nested RT-PCR.

Conclusions/interpretation: Our data support the concept that within the exocrine (acinar and ductal) pancreas of the adult mouse there are cells that can give rise to insulin-positive cells in vitro.

Citing Articles

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β-cell replacement sources for type 1 diabetes: a focus on pancreatic ductal cells.

Corritore E, Lee Y, Sokal E, Lysy P Ther Adv Endocrinol Metab. 2016; 7(4):182-99.

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Intraislet Pancreatic Ducts Can Give Rise to Insulin-Positive Cells.

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Li D, Peng S, Zhang Z, Feng R, Li L, Liang J J Zhejiang Univ Sci B. 2013; 14(7):596-603.

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Purified human pancreatic duct cell culture conditions defined by serum-free high-content growth factor screening.

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