Generation and Characterization of Human Cryptorchid-Specific Induced Pluripotent Stem Cells from Urine
Generation and Characterization of Human Cryptorchid-Specific Induced Pluripotent Stem Cells from Urine
复制标题
从尿液中产生人类隐睾特异性诱导多能干细胞
DOI：
10.1089/scd.2012.0260
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发表时间：
2013-03-01
影响因子：
4
通讯作者：
Chen, Fang
中科院分区：
文献类型：
--
作者：
Zhou, Junmei;Wang, Xue;Chen, Fang
隐睾症是人类常见的先天性出生缺陷，可能并发不孕症。由于人类胚胎无法用于研究目的，隐睾的体外模型可能是有价值的。在这项研究中，我们重新编程尿细胞含有胰岛素样因子3，锌指（ZNF）214，和ZNF 215的遗传变异隐睾患者通过引入人类OCT 4，SOX 2，C-MYC，和KLF 4与慢病毒。然后将细胞重新铺在经辐照的小鼠胚胎成纤维细胞上，并与人胚胎干（ES）细胞培养基一起培养。手动挑取具有明确边界的紧密集落，并对2种诱导多能细胞系进行充分表征。我们的研究结果表明，这2个细胞系在形态学外观，标记物表达和多能细胞特异性基因OCT 4的表观遗传状态方面与人ES细胞相似。这些细胞在畸胎瘤和体外可分化为所有3个胚层的细胞，包括分化为VASA阳性生殖细胞谱系。亲本尿细胞和重编程细胞均具有正常的核型和相同的短串联重复序列位点，表明这两个细胞群体具有相同的遗传特性。本研究成功构建了人尿源性隐睾特异性诱导多能干细胞，为隐睾发病机制的研究提供了良好的遗传学基础。
Cryptorchidism is a common congenital birth defect in human beings with the possible complication of infertility. An in vitro model of cryptorchidism might be valuable due to the inaccessibility of human embryos for research purposes. In this study, we reprogrammed urine cells containing genetic variations in insulin-like factor 3, zinc finger (ZNF) 214, and ZNF215 from a cryptorchid patient by introducing human OCT4, SOX2, C-MYC, and KLF4 with lentivirus. The cells were then replated on irradiated mouse embryonic fibroblasts and cultured with the human embryonic stem (ES) cell medium. The compact colonies with well-defined borders were manually picked, and 2 induced pluripotent cell lines were fully characterized. Our results demonstrated that these 2 cell lines were similar to human ES cells in morphological appearance, marker expression, and epigenetic status of the pluripotent cell-specific gene, OCT4. These cells could be differentiated into cells of all 3 germ layers in teratomas and in vitro, including into the VASA-positive germ cell lineage. Both parental urine cells and the reprogrammed cells possessed the normal karyotype and the same short tandem repeat loci, indicating that these 2 cell population share the same genetic identity. This establishment and characterization of human urine-derived cryptorchid-specific induced pluripotent stem cells could present a good human genetic system for future studies investigating the molecular mechanism of cryptorchidism.