Analysis of Insulin-Producing Cells During In Vitro Differentiation From Feeder-Free Embryonic Stem Cells

  • Yusuke Moritoh
    From the Division of Stem Cell Regulation Research, Osaka University Graduate School of Medicine, Suita, Japan
  • Eiji Yamato
    From the Division of Stem Cell Regulation Research, Osaka University Graduate School of Medicine, Suita, Japan
  • Yumiko Yasui
    From the Division of Stem Cell Regulation Research, Osaka University Graduate School of Medicine, Suita, Japan
  • Satsuki Miyazaki
    From the Division of Stem Cell Regulation Research, Osaka University Graduate School of Medicine, Suita, Japan
  • Jun-ichi Miyazaki
    From the Division of Stem Cell Regulation Research, Osaka University Graduate School of Medicine, Suita, Japan

抄録

<jats:p>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.</jats:p>

収録刊行物

  • Diabetes

    Diabetes 52 (5), 1163-1168, 2003-05-01

    American Diabetes Association

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