Regulated expression of pdx-1 promotes in vitro differentiation of insulin-producing cells from embryonic stem cells
Regulated expression of pdx-1 promotes in vitro differentiation of insulin-producing cells from embryonic stem cells
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DOI：
10.2337/diabetes.53.4.1030
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发表时间：
2004-04-01
期刊：
影响因子：
7.7
通讯作者：
Miyazaki, J
中科院分区：
文献类型：
--
作者：
Miyazaki, S;Yamato, E;Miyazaki, J
胚胎干细胞可以分化为多种类型的细胞。最近的报道表明，ES细胞可以分化为产生胰岛素的细胞。然而，分化效率不足以产生胰岛素分泌细胞用于未来的治疗。PDX-1是一种含有同源结构域的转录因子，是胰腺发育的重要调节因子。我们建立了一个外源PDX-1表达受整合到rosa26基因座上的Tet-off系统精确调控的ES细胞系。利用该细胞系，我们检测了PDX-1在体外分化过程中通过类胚体形成对PDX-1表达的影响。结果表明，PDX-1的表达明显增强了分化细胞中胰岛素2、生长抑素、Kir6.2、葡萄糖激酶、神经原蛋白3、P48、Pax6、PC2和HNF6基因的表达。免疫组织化学检测还显示，大多数分化的ES细胞都有高水平的胰岛素产生。因此，外源表达PDX-1将为有效地从人ES细胞中产生胰岛素分泌细胞提供一种有前景的方法，用于未来糖尿病患者的治疗。
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.