Monitoring and robust induction of nephrogenic intermediate mesoderm from human pluripotent stem cells.

Monitoring and robust induction of nephrogenic intermediate mesoderm from human pluripotent stem cells.
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DOI:
10.1038/ncomms2378
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发表时间:
2013
影响因子:
16.6
通讯作者:
--
中科院分区:
综合性期刊1区
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用于刺激人多能干细胞(hPSC)分化成肾谱系的方法仍有待开发。肾脏中的大多数细胞来源于称为中间中胚层(IM)的胚胎胚层。在这里,我们展示了通过基于细菌人工染色体(BAC)的载体和基于单核苷酸多态性(SNP)阵列的检测在hPSC中建立有效的同源重组系统。该系统使我们能够产生含有敲入OSR 1(IM的特异性标记物)的绿色荧光蛋白(GFP)的人诱导多能干细胞(hiPSC)系。我们还建立了一个强大的IM诱导方案,可产生高达90%的OSR 1+细胞。这些人IM细胞可以在体外和体内分化成IM衍生器官的多种细胞类型,从而提供了一个前所未有的系统来阐明IM发育的机制,并可能为肾脏的再生治疗提供细胞来源。
A method for stimulating the differentiation of human pluripotent stem cells (hPSCs) into kidney lineages remains to be developed. Most cells in kidney are derived from an embryonic germ layer known as intermediate mesoderm (IM). Here we show the establishment of an efficient system of homologous recombination in hPSCs by means of bacterial artificial chromosome (BAC)-based vectors and single nucleotide polymorphism (SNP) array-based detection. This system allowed us to generate human induced pluripotent stem cell (hiPSC) lines containing green fluorescence protein (GFP) knocked into OSR1, a specific marker for IM. We have also established a robust induction protocol for IM, which produces up to 90% OSR1+ cells. These human IM cells can differentiate into multiple cell types of IM-derived organs in vitro and in vivo, thereby supplying an unprecedented system to elucidate the mechanisms of IM development and potentially providing a cell source for regenerative therapies of the kidney.
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