Branching ducts similar to mesonephric ducts or ureteric buds in teratomas originating from mouse embryonic stem cells
Branching ducts similar to mesonephric ducts or ureteric buds in teratomas originating from mouse embryonic stem cells
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DOI：
10.1152/ajprenal.00001.2004
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发表时间：
2006-01-01
影响因子：
4.2
通讯作者：
Sasaki, K
中科院分区：
文献类型：
--
作者：
Yamamoto, M;Cui, L;Sasaki, K
分支管类似于源自小鼠胚胎干细胞的畸胎瘤中的中肾管或输尿管芽。 Am J Physiol Renal Physiol 290: F52 - F60, 2006。首次发表于 2005 年 8 月 16 日； doi:10.1152/ajprenal.00001.2004.- 构成后肾肾原基的输尿管芽上皮细胞和后肾间充质细胞能够通过相互诱导相互作用产生肾单位和集合管。一旦这些细胞由多能胚胎干（ES）细胞诱导产生，它们就有可能成为肾组织再生的强大工具。在这项研究中，我们研究了小鼠 ES 细胞生长物及其移植物中的这些肾原始细胞和结构。通过 RT-PCR 在成年小鼠的胚状体 (EB) 生长物及其移植物中检查早期肾脏发育必需的基因表达。在移植组织中进行肾脏原始结构的组织化学检测和基因表达分析以及激光显微切割。 RT-PCR 分析检测了扩增 6-9 天后的 EB 生长物中 Pax-2、Lim-1、c-Ret、Emx2、Sall1、WT-1、Eya-1、GDNF 和 Wnt-4 的基因表达，以及移植后 14 和 28 天的畸胎瘤组织中的基因表达。移植后 14 天的组织化学分析显示，一些导管对 Pax-2、endo A 细胞角蛋白、肾脏特异性钙粘蛋白和 Dolichos biflorus 凝集素呈阳性，并且这些导管已发生二叉分支。这些染色模式和形态特征是中肾管和输尿管芽固有的。在28天的长期存活中，Pax-2免疫反应性在一些肾原基样结构中消失，表明它们的分化。一些导管伴有中肾肾单位样曲小管。对显微切割收集的这些结构进行 RT-PCR 分析证实，它们表达与肾脏发育相关的基因。总之，这些数据表明 ES 细胞具有产生肾原始管结构的潜力，并提供了对肾组织再生的深入了解。
Branching ducts similar to mesonephric ducts or ureteric buds in teratomas originating from mouse embryonic stem cells. Am J Physiol Renal Physiol 290: F52 - F60, 2006. First published August 16, 2005; doi:10.1152/ajprenal.00001.2004.- Ureteric bud epithelial cells and metanephric mesenchymal cells that comprise the metanephric kidney primordium are capable of producing nephrons and collecting ducts through reciprocal inductive interaction. Once these cells are induced from pluripotent embryonic stem (ES) cells, they have the potential to become powerful tools in the regeneration of kidney tissues. In this study, we investigated these renal primordial cells and structures in mouse ES cell outgrowths and their transplants. Gene expression essential for early kidney development was examined by RT-PCR in embryoid body (EB) outgrowths and their transplants in adult mice. Histochemical detection of kidney primordial structures and gene expression analysis coupled with laser microdissection were performed in transplant tissues. RT-PCR analysis detected gene expression of Pax-2, Lim-1, c-Ret, Emx2, Sall1, WT-1, Eya-1, GDNF, and Wnt-4 in the EB outgrowths from days 6 - 9 of expansion onward, and also in the teratoma tissues 14 and 28 days after transplantation. Histochemical analysis 14 days after transplantation showed that some ducts were positive for Pax-2, endo A cytokeratin, kidney-specific cadherin, and Dolichos biflorus agglutinin and that dichotomous branching of these ducts had occurred. These staining patterns and morphological features are intrinsic for mesonephric ducts and ureteric buds. In long-term survival of 28 days, Pax-2-immunoreactivity disappeared in some renal primordia-like structures, indicating their differentiation. Some ducts were accompanied by mesonephric nephron-like convoluted tubules. RT-PCR analysis of those structures collected by microdissection confirmed that they expressed kidney development-related genes. In conclusion, these data suggest the potential of ES cells to produce renal primordial duct structures and provides an insight into the regeneration of kidney tissues.