Establishment of canine iPS cells for the study of disease-model dogs

为了建立犬iPS细胞，利用EOS载体系统将犬4重编程因子导入犬胚胎成纤维细胞，通过耐药性和GFP荧光检测筛选出内源性Oct 3/4基因表达的克隆。选择的克隆是阳性的干细胞标记基因，并能够使胚状体，这表明诱导的标记基因特异性的三个胚层。 然而，未观察到畸胎瘤形成。 使用化学成分确定的培养基，这些克隆在几代中保持未分化状态，但长期培养迫使这些克隆自发分化，因此没有建立具有与iPS细胞相容的特征的克隆。 这些数据表明，将需要更多的研究，以找到适合犬iPS细胞的培养条件。

To establish canine iPS cells, canine four reprogramming factor were introduced into canine embryonic fibroblasts with the EOS vector system, which allowed us to select clones with the expression of endogenous Oct3/4 gene by drug-resistance and fluorescence of GFP. Selected clones were positive for stem cell marker genes and able to make embryoid bodies, which showed the induction of marker genes specific for each of three germ layers. Teratoma formation was, however, not observed. Using chemically defined medium, these clones were maintained at undifferentiated state during several passages, but the long term culture forced the spontaneous differentiation of these clones, and therefore no clones with the feature compatible with iPS cells were established. These data indicated that more investigation will be required to find the suitable culture condition for canine iPS cells.