Saitoh Koichi, Yamato Eiji, Miyazaki Satsuki, Miyazaki Jun-Ichi
Division of Stem Cell Regulation Research, Osaka University Graduate School of Medicine, Osaka, Japan.
Diabetes Res Clin Pract. 2007 Sep;77 Suppl 1:S138-42. doi: 10.1016/j.diabres.2007.01.048. Epub 2007 Apr 23.
Embryonic stem (ES) cells can differentiate into many cell types. Recent reports have shown that ES cells can differentiate into insulin-producing cells. We have established an ES cell line in which exogenous Pdx-1 expression was precisely regulated by the Tet-off system integrated into the ROSA26 locus and succeeded to produce insulin-producing cells. The Pdx-1 expressing final differentiated insulin-positive cells can be maintained for more than 2 months. However, in spite of their induced expression of Pdx-1, the repeated passages of cells lost their capacity to express insulin and NeuroD1 gene. Forced expression of NeuroD1 gene by adenoviral vector in these cells restored the expression of insulin. These results suggested that maintenance of the property of insulin-producing cells derived from ES cells could be achieved by synergistic expression of Pdx-1 and NeuroD1.
胚胎干细胞（ES细胞）可分化为多种细胞类型。最近的报道表明，ES细胞可分化为产生胰岛素的细胞。我们建立了一种ES细胞系，其中整合到ROSA26位点的Tet-off系统精确调控外源Pdx-1的表达，并成功产生了产生胰岛素的细胞。表达Pdx-1的最终分化胰岛素阳性细胞可维持2个月以上。然而，尽管这些细胞诱导表达了Pdx-1，但细胞的反复传代使其失去了表达胰岛素和NeuroD1基因的能力。通过腺病毒载体在这些细胞中强制表达NeuroD1基因可恢复胰岛素的表达。这些结果表明，通过Pdx-1和NeuroD1的协同表达可实现对ES细胞来源的产生胰岛素细胞特性的维持。
