In vitro derivation of functional insulin-producing cells from human embryonic stem cells
In vitro derivation of functional insulin-producing cells from human embryonic stem cells
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DOI：
10.1038/cr.2007.28
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发表时间：
2007-04-01
期刊：
影响因子：
44.1
通讯作者：
Deng, Hongkui
中科院分区：
文献类型：
--
作者：
Jiang, Wei;Shi, Yan;Deng, Hongkui
人胚胎干细胞（ES）的自我更新和分化能力使其成为产生用于治疗I型糖尿病的胰腺β细胞的潜在来源。在这里，我们报告了一种新开发的和有效的方法，在无血清系统中进行，诱导人ES细胞分化为胰岛素产生细胞。激活素A在初始阶段用于诱导人ES细胞的定形内胚层分化，如通过定形内胚层标志物Sox 17和Brachyury的表达所检测的。此外，全反式视黄酸（RA）用于促进胰腺分化，如早期胰腺转录因子pdx 1和hlxb 9的表达所示。在含有bFGF和烟酰胺的DMEM/F12无血清培养基中成熟后，分化的细胞表达胰岛特异性标志物，如C肽、胰岛素、胰高血糖素和β 2。C肽阳性细胞百分比超过15%。这些细胞分泌的胰岛素和C肽与葡萄糖水平的变化相对应。当移植到链脲佐菌素（STZ）处理的裸鼠的肾包膜中时，这些分化的人ES细胞存活并保持β细胞标记基因的表达，包括C肽，pdx 1，葡萄糖激酶，nkx 6.1，IAPP，pax 6和7 cf 1。30%的移植裸鼠表现出明显的稳定性恢复。并且校正的表型持续超过六周。我们的新方法为研究人胰腺发育机制提供了一个有前途的体外分化模型，并说明了使用人ES细胞治疗I型糖尿病的潜力。
The capacity for self-renewal and differentiation of human embryonic stem (ES) cells makes them a potential source for generation of pancreatic beta cells for treating type I diabetes mellitus. Here, we report a newly developed and effective method, carried out in a serum-free system, which induced human ES cells to differentiate into insulin-producing cells. Activin A was used in the initial stage to induce definitive endoderm differentiation from human ES cells, as detected by the expression of the definitive endoderm markers Sox17 and Brachyury. Further, all-trans retinoic acid (RA) was used to promote pancreatic differentiation, as indicated by the expression of the early pancreatic transcription factors pdx1 and hlxb9. After maturation in DMEM/F12 serum-free medium with bFGF and nicotinamide, the differentiated cells expressed islet specific markers such as C-peptide, insulin, glucagon and glut2. The percentage of C-peptide-positive cells exceeded 15%. The secretion of insulin and C-peptide by these cells corresponded to the variations in glucose levels. When transplanted into renal capsules of Streptozotocin (STZ)-treated nude mice, these differentiated human ES cells survived and maintained the expression of beta cell marker genes, including C-peptide, pdx1, glucokinase, nkx6.1, IAPP, pax6 and 7cf1. Thirty percent of the transplanted nude mice exhibited apparent restoration of stable euglycemia.; and the corrected phenotype was sustained for more than six weeks. Our new method provides a promising in vitro differentiation model for studying the mechanisms of human pancreas development and illustrates the potential of using human ES cells for the treatment of type I diabetes mellitus.