Generation of Induced Pluripotent Stem Cells in Rabbits POTENTIAL EXPERIMENTAL MODELS FOR HUMAN REGENERATIVE MEDICINE
Generation of Induced Pluripotent Stem Cells in Rabbits POTENTIAL EXPERIMENTAL MODELS FOR HUMAN REGENERATIVE MEDICINE
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DOI：
10.1074/jbc.m110.150540
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发表时间：
2010-10-08
影响因子：
4.8
通讯作者：
Ogura, Atsuo
中科院分区：
文献类型：
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作者：
Honda, Arata;Hirose, Michiko;Ogura, Atsuo
人类诱导多能干（iPS）细胞有潜力建立一个有前景的再生医学新领域。因此，在开始人体试验之前，必须使用适当的动物模型严格测试 iPS 衍生细胞的安全性和效率。在这里，我们报告了兔 iPS 细胞的建立，这是第一个由小型实验动物物种产生的人型 iPS 细胞。使用慢病毒载体，将四种人类重编程基因（c-MYC、KLF4、SOX2 和 OCT3/4）成功导入成年兔肝细胞和胃细胞中。由此产生的兔 iPS 细胞与人类 iPS 细胞非常相似。它们形成具有锋利边缘的扁平集落，并在碱性 FGF 存在下无限增殖。它们表达内源性多能性标记物 c-MYC、KLF4、SOX2、OCT3/4 和 NANOG，而引入的人类基因则完全沉默。使用体外分化条件，兔 iPS 细胞很容易分化为外胚层、中胚层和内胚层。他们还在免疫缺陷小鼠体内形成了含有所有三个胚层的多种组织的畸胎瘤。因此，兔 iPS 细胞满足了获得完全重编程状态的所有要求，与我们最近生成的胚胎干细胞对应物表现出高度相似性。然而，他们的全局基因表达分析揭示了这两种类型的兔多能干细胞之间存在轻微但严格的差异。兔子模型应该使我们能够在相同的标准化条件下比较 iPS 细胞和胚胎干细胞，以评估它们在人类中基于多能细胞的再生医学的最终可行性。
Human induced pluripotent stem (iPS) cells have the potential to establish a new field of promising regenerative medicine. Therefore, the safety and the efficiency of iPS-derived cells must be tested rigorously using appropriate animal models before human trials can commence. Here, we report the establishment of rabbit iPS cells as the first human-type iPS cells generated from a small laboratory animal species. Using lentiviral vectors, four human reprogramming genes (c-MYC, KLF4, SOX2, and OCT3/4) were introduced successfully into adult rabbit liver and stomach cells. The resulting rabbit iPS cells closely resembled human iPS cells; they formed flattened colonies with sharp edges and proliferated indefinitely in the presence of basic FGF. They expressed the endogenous pluripotency markers c-MYC, KLF4, SOX2, OCT3/4, and NANOG, whereas the introduced human genes were completely silenced. Using in vitro differentiating conditions, rabbit iPS cells readily differentiated into ectoderm, mesoderm, and endoderm. They also formed teratomas containing a variety of tissues of all three germ layers in immunodeficient mice. Thus, the rabbit iPS cells fulfilled all of the requirements for the acquisition of the fully reprogrammed state, showing high similarity to their embryonic stem cell counterparts we generated recently. However, their global gene expression analysis revealed a slight but rigid difference between these two types of rabbit pluripotent stem cells. The rabbit model should enable us to compare iPS cells and embryonic stem cells under the same standardized conditions in evaluating their ultimate feasibility for pluripotent cell-based regenerative medicine in humans.