Derivation and Characterization of Induced Pluripotent Stem Cells from Equine Fibroblasts
Derivation and Characterization of Induced Pluripotent Stem Cells from Equine Fibroblasts
复制标题
DOI：
10.1089/scd.2012.0052
复制
发表时间：
2013-02-01
影响因子：
4
通讯作者：
Donadeu, Francesc Xavier
中科院分区：
文献类型：
--
作者：
Breton, Amandine;Sharma, Ruchi;Donadeu, Francesc Xavier
多能干细胞不仅为人类医学提供了前所未有的潜力，而且也为兽医学，特别是与马有关的医学提供了前所未有的潜力。诱导多能干细胞（iPSCs）特别有前景，因为它们在功能上与胚胎干细胞相似，并且可以在体外以患者特异性的方式生成。在这项研究中，我们报道了从马驹身上获得的皮肤成纤维细胞产生马iPSCs，并使用编码小鼠Oct4， Sox2， c-Myc和Klf4序列的病毒载体重新编程。重编程的细胞系在形态上与其他物种的iPSCs相似，并且可以在30多次传代中稳定维持。免疫染色和聚合酶链反应分析显示，这些细胞系表达了一系列与多能性相关的内源性标记，包括OCT4、SOX2、NANOG、REX1、LIN28、SSEA1、SSEA4和TRA1-60。此外，在适当的条件下，马iPSCs容易形成胚状体，并在体外分化为表达外胚层、中胚层和内胚层标记的细胞，注射到免疫缺陷小鼠体内后，产生含有这3种胚层分化衍生物的肿瘤。最后，我们还重新编程了一匹2岁马的成纤维细胞。重编程细胞在形态、多能性标记物的表达和分化能力方面与来自新生儿成纤维细胞的iPSCs相似。这些新细胞系的产生是了解马多能性的重要一步，并为iPSC技术潜在地成为兽医生物医学强有力的研究和临床工具铺平了道路。
Pluripotent stem cells offer unprecedented potential not only for human medicine but also for veterinary medicine, particularly in relation to the horse. Induced pluripotent stem cells (iPSCs) are particularly promising, as they are functionally similar to embryonic stem cells and can be generated in vitro in a patient-specific manner. In this study, we report the generation of equine iPSCs from skin fibroblasts obtained from a foal and reprogrammed using viral vectors coding for murine Oct4, Sox2, c-Myc, and Klf4 sequences. The reprogrammed cell lines were morphologically similar to iPSCs reported from other species and could be stably maintained over more than 30 passages. Immunostaining and polymerase chain reaction analyses revealed that these cell lines expressed an array of endogenous markers associated with pluripotency, including OCT4, SOX2, NANOG, REX1, LIN28, SSEA1, SSEA4, and TRA1-60. Furthermore, under the appropriate conditions, the equine iPSCs readily formed embryoid bodies and differentiated in vitro into cells expressing markers of ectoderm, mesoderm, and endoderm, and when injected into immunodeficient mice, gave raise to tumors containing differentiated derivatives of the 3 germ layers. Finally, we also reprogrammed fibroblasts from a 2-year-old horse. The reprogrammed cells were similar to iPSCs derived from neonatal fibroblasts in terms of morphology, expression of pluripotency markers, and differentiation ability. The generation of these novel cell lines constitutes an important step toward the understanding of pluripotency in the horse, and paves the way for iPSC technology to potentially become a powerful research and clinical tool in veterinary biomedicine.