Department of Animal Science, Michigan State University, East Lansing, Michigan 48824, USA.
Stem Cells Dev. 2011 Oct;20(10):1669-78. doi: 10.1089/scd.2011.0127. Epub 2011 Jun 15.
For more than thirty years, the dog has been used as a model for human diseases. Despite efforts made to develop canine embryonic stem cells, success has been elusive. Here, we report the generation of canine induced pluripotent stem cells (ciPSCs) from canine adult fibroblasts, which we accomplished by introducing human OCT4, SOX2, c-MYC, and KLF4. The ciPSCs expressed critical pluripotency markers and showed evidence of silencing the viral vectors and normal karyotypes. Microsatellite analysis indicated that the ciPSCs showed the same profile as the donor fibroblasts but differed from cells taken from other dogs. Under culture conditions favoring differentiation, the ciPSCs could form cell derivatives from the ectoderm, mesoderm, and endoderm. Further, the ciPSCs required leukemia inhibitory factor and basic fibroblast growth factor to survive, proliferate, and maintain pluripotency. Our results demonstrate an efficient method for deriving canine pluripotent stem cells, providing a powerful platform for the development of new models for regenerative medicine, as well as for the study of the onset, progression, and treatment of human and canine genetic diseases.
三十多年来，狗一直被用作人类疾病的模型。尽管人们努力开发犬类胚胎干细胞，但一直未能成功。在这里，我们报告了从犬类成纤维细胞中生成犬类诱导多能干细胞（ciPSCs）的方法，我们通过引入人 OCT4、SOX2、c-MYC 和 KLF4 来实现这一目标。ciPSCs 表达了关键的多能性标记，并显示出沉默病毒载体和正常核型的证据。微卫星分析表明，ciPSCs 与供体细胞的表现相同，但与来自其他犬只的细胞不同。在有利于分化的培养条件下，ciPSCs 可以从外胚层、中胚层和内胚层形成细胞衍生物。此外，ciPSCs 需要白血病抑制因子和碱性成纤维细胞生长因子才能存活、增殖并维持多能性。我们的结果证明了一种高效的犬类多能干细胞的衍生方法，为开发新的再生医学模型以及研究人类和犬类遗传疾病的发病、进展和治疗提供了强大的平台。
