Miyazaki Satsuki, Yamato Eiji, Miyazaki Jun-ichi
Division of Stem Cell Regulation Research, Osaka University Graduate School of Medicine, Suita, Japan.
Diabetes. 2004 Apr;53(4):1030-7. doi: 10.2337/diabetes.53.4.1030.
Embryonic stem (ES) cells can differentiate into many cell types. Recent reports have shown that ES cells can differentiate into insulin-producing cells. However, the differentiation is not efficient enough to produce insulin-secreting cells for future therapeutic use. Pdx-1, a homeodomain-containing transcription factor, is a crucial regulator for pancreatic development. We established an ES cell line in which exogenous pdx-1 expression was precisely regulated by the Tet-off system integrated into the ROSA26 locus. Using this cell line, we examined the effect of pdx-1 expression during in vitro differentiation via embryoid body formation. The results showed that pdx-1 expression clearly enhanced the expression of the insulin 2, somatostatin, Kir6.2, glucokinase, neurogenin3, p48, Pax6, PC2, and HNF6 genes in the resulting differentiated cells. Immunohistochemical examination also revealed that insulin was highly produced in most of the differentiated ES cells. Thus, exogenous expression of pdx-1 should provide a promising approach for efficiently producing insulin-secreting cells from human ES cells for future therapeutic use in diabetic patients.
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。然而，这种分化效率不足以产生用于未来治疗的胰岛素分泌细胞。Pdx-1是一种含同源结构域的转录因子，是胰腺发育的关键调节因子。我们建立了一种ES细胞系，其中外源pdx-1的表达由整合到ROSA26位点的Tet-off系统精确调控。利用该细胞系，我们通过胚状体形成研究了体外分化过程中pdx-1表达的影响。结果表明，pdx-1的表达明显增强了所得分化细胞中胰岛素2、生长抑素、Kir6.2、葡萄糖激酶、神经生成素3、p48、Pax6、PC2和HNF6基因的表达。免疫组织化学检查还显示，大多数分化的ES细胞中大量产生胰岛素。因此，外源表达pdx-1应为从人ES细胞高效产生胰岛素分泌细胞用于未来糖尿病患者的治疗提供一种有前景的方法。
