Fundamental investigation on the cell transplantation therapy to the chronic renal failure using the feline iPS cell

采用AGPC法从猫组织中提取总rna。利用Oligo-dT乳胶珠法将总rna纯化为mRNA，成功克隆了猫Klf4、Oct3/4、Sox2和AQP2基因。将猫Klf4、Oct3/4和Sox2的CDS插入pCl-neo哺乳动物载体中。从猫皮肤中获得的成纤维细胞尝试了这些载体的共转染，但iPS细胞不能诱导。将猫AQP2基因插入到pTrancer-EF-Bsd中，将该载体转染到CrFK细胞系和293细胞系。利用该细胞进行吸水研究，能分离出少量高水平表达GFP的细胞株，吸水率的增加明显高于对照CrFK细胞株。

Total-RNA was extracted from a feline tissue by using the AGPC method. Further total-RNA was purified to mRNA by the Oligo-dT latex beads method, and using this mRNA, the cloning of feline Klf4, Oct3/4, Sox2 and AQP2 gene was successful. The CDS of feline Klf4, Oct3/4 and Sox2 were inserted into pCl-neo Mammalian Vector. The fibroblast which it obtained from a feline skin tried the co-transfection of these vectors, but the iPS cell was not able to induce. While, feline AQP2 gene was inserted into pTrancer-EF-Bsd, and this vector was transfected into CrFK cell line and 293 cells line. It could isolate few high level expression GFP cell lines, and the increase of the water absorption was significantly higher than control CrFK cell line in the water absorption study using the cell.