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

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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.