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Experimental Diabetes Research
Volume 2012 (2012), Article ID 201295, 5 pages
http://dx.doi.org/10.1155/2012/201295
Methodology Report

Efficient Differentiation of Mouse Embryonic Stem Cells into Insulin-Producing Cells

Pharmacognosy Lab, Herbal Medicinal Product Technology Division, Industrial Technology Research Institute, Hsinchu 30011, Taiwan

Received 23 February 2012; Revised 22 June 2012; Accepted 5 July 2012

Academic Editor: Joseph R. Landolph Jr.

Copyright © 2012 Szu-Hsiu Liu and Lain-Tze Lee. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Embryonic stem (ES) cells are a potential source of a variety of differentiated cells for cell therapy, drug discovery, and toxicology screening. Here, we present an efficacy strategy for the differentiation of mouse ES cells into insulin-producing cells (IPCs) by a two-step differentiation protocol comprising of (i) the formation of definitive endoderm in monolayer culture by activin A, and (ii) this monolayer endoderm being induced to differentiate into IPCs by nicotinamide, insulin, and laminin. Differentiated cells can be obtained within approximately 7 days. The differentiation IPCs combined application of RT-PCR, ELISA, and immunofluorescence to characterize phenotypic and functional properties. In our study, we demonstrated that IPCs produced pancreatic transcription factors, endocrine progenitor marker, definitive endoderm, pancreatic β-cell markers, and Langerhans α and δ cells. The IPCs released insulin in a manner that was dose dependent upon the amount of glucose added. These techniques may be able to be applied to human ES cells, which would have very important ramifications for treating human disease.