PT  - JOURNAL ARTICLE
AU  - Moritoh, Yusuke
AU  - Yamato, Eiji
AU  - Yasui, Yumiko
AU  - Miyazaki, Satsuki
AU  - Miyazaki, Jun-ichi
TI  - Analysis of Insulin-Producing Cells During In Vitro Differentiation From Feeder-Free Embryonic Stem Cells
AID  - 10.2337/diabetes.52.5.1163
DP  - 2003 May 01
TA  - Diabetes
PG  - 1163--1168
VI  - 52
IP  - 5
4099  - http://diabetes.diabetesjournals.org/content/52/5/1163.short
4100  - http://diabetes.diabetesjournals.org/content/52/5/1163.full
SO  - Diabetes2003 May 01; 52
AB  - Embryonic stem (ES) cells can differentiate into many cell types and are expected to be useful for tissue engineering. Recent reports have shown that ES cells can differentiate into insulin-producing cells in response to the transient expression of the pdx-1 gene, after the removal of feeder cells. To investigate the lineage of insulin-producing cells and their in vitro differentiation, we introduced the βgeo gene, encoding a β-galactosidase-neomycin phosphotransferase fusion protein under the control of the mouse insulin 2 promoter, into ES cells that had been adapted to feeder-free culture, and analyzed insulin gene expression during their in vitro differentiation. We also examined the expression of transcription factors that are related to the differentiation of the pancreas. X-gal staining analysis revealed β-galactosidase-positive cells on the surface and in the center of the embryoid body that proliferated during differentiation. Glucose-responsive insulin-producing cells, derived from our feeder-free ES cells, expressed insulin 2, pdx-1, Pax4, and Isl1 and also the glucagon, somatostatin, and PP genes. Moreover, the genes encoding p48, amylase, and carboxypeptidase A were also expressed. These results suggest that ES cells can differentiate not only into endocrine cells but also into exocrine cells of the pancreas, without the initiation of pdx-1 expression.