Colony-forming cells in the adult mouse pancreas are expandable in Matrigel and form endocrine/acinar colonies in laminin hydrogel

Item Type:	Article
Title:	Colony-forming cells in the adult mouse pancreas are expandable in Matrigel and form endocrine/acinar colonies in laminin hydrogel
Creators Name:	Jin, L. and Feng, T. and Shih, H.P. and Zerda, R. and Luo, A. and Hsu, J. and Mahdavi, A. and Sander, M. and Tirrell, D.A. and Riggs, A.D. and Ku, H.T.
Abstract:	The study of hematopoietic colony-forming units using semisolid culture media has greatly advanced the knowledge of hematopoiesis. Here we report that similar methods can be used to study pancreatic colony-forming units. We have developed two pancreatic colony assays that enable quantitative and functional analyses of progenitor-like cells isolated from dissociated adult (2-4 mo old) murine pancreas. We find that a methylcellulose-based semisolid medium containing Matrigel allows growth of duct-like "Ring/Dense" colonies from a rare (∼1%) population of total pancreatic single cells. With the addition of roof plate-specific spondin 1, a wingless-int agonist, Ring/Dense colony-forming cells can be expanded more than 100,000-fold when serially dissociated and replated in the presence of Matrigel. When cells grown in Matrigel are then transferred to a Matrigel-free semisolid medium with a unique laminin-based hydrogel, some cells grow and differentiate into another type of colony, which we name "Endocrine/Acinar." These Endocrine/Acinar colonies are comprised mostly of endocrine- and acinar-like cells, as ascertained by RNA expression analysis, immunohistochemistry, and electron microscopy. Most Endocrine/Acinar colonies contain beta-like cells that secrete insulin/C-peptide in response to D-glucose and theophylline. These results demonstrate robust self-renewal and differentiation of adult Ring/Dense colony-forming units in vitro and suggest an approach to producing beta-like cells for cell replacement of type 1 diabetes. The methods described, which include microfluidic expression analysis of single cells and colonies, should also advance study of pancreas development and pancreatic progenitor cells.
Keywords:	Extracellular Matrix Proteins, Sry-related HMG Box (Sox) 9, Promonin 1(CD133), Neurogenin 3, Dickkopf1 (Dkk1), Animals, Mice
Source:	Proceedings of the National Academy of Sciences of the United States of America
ISSN:	1091-6490
Publisher:	National Academy of Sciences
Volume:	110
Number:	10
Page Range:	3907-12
Date:	5 March 2013
Official Publication:	https://doi.org/10.1073/pnas.1301889110
PubMed:	View item in PubMed

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