Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
Cancer Genomics Project, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan.
Nat Commun. 2013;4:1367. doi: 10.1038/ncomms2378.
A method for stimulating the differentiation of human pluripotent stem cells into kidney lineages remains to be developed. Most cells in kidney are derived from an embryonic germ layer known as intermediate mesoderm. Here we show the establishment of an efficient system of homologous recombination in human pluripotent stem cells by means of bacterial artificial chromosome-based vectors and single-nucleotide polymorphism array-based detection. This system allowed us to generate human-induced pluripotent stem cell lines containing green fluorescence protein knocked into OSR1, a specific intermediate mesoderm marker. We have also established a robust induction protocol for intermediate mesoderm, which produces up to 90% OSR1(+) cells. These human intermediate mesoderm cells can differentiate into multiple cell types of intermediate mesoderm-derived organs in vitro and in vivo, thereby supplying a useful system to elucidate the mechanisms of intermediate mesoderm development and potentially providing a cell source for regenerative therapies of the kidney.
一种将人类多能干细胞诱导分化为肾系细胞的方法仍有待开发。肾脏中的大多数细胞来源于一种称为中胚层的胚胎生殖层。在这里，我们通过基于细菌人工染色体的载体和基于单核苷酸多态性阵列的检测，展示了在人类多能干细胞中建立同源重组的有效系统。该系统使我们能够生成含有绿色荧光蛋白的人诱导多能干细胞系，该蛋白敲入特定的中胚层标记物 OSR1。我们还建立了一种强大的中胚层诱导方案，可产生高达 90%的 OSR1(+)细胞。这些人类中胚层细胞可以在体外和体内分化为多种中胚层来源的器官细胞类型，从而提供了一个有用的系统来阐明中胚层发育的机制，并可能为肾脏的再生治疗提供细胞来源。
