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. 2014 Feb 20;23(13):1515–1523. doi: 10.1089/scd.2013.0461

FIG. 1.

FIG. 1.

EquiPSCs exhibit a characteristic pluripotent stem cell morphology and express pluripotency factors. (A) At passage 1, equiPSCs have a characteristic pluripotent stem cell morphology with a high nuclear-to-cytoplasmic ratio. (B) Colonies have a compact, domed appearance. (C) EquiPSCs have a normal 64,XX karyotype. (D) At passage 6, all seven of the selected clones showed robust expression of the four transgenes as demonstrated in this figure with clone K. At passage 25, clones H, I, and K retained expression of the hOCT4 transgene, while Clone H also continued to express hSOX2. (E) EquiPSCs are strongly positive for alkaline phosphatase activity. (F) RT-PCR expression analysis of equine pluripotency factors NANOG, OCT4, and TERT, and positive control β-ACTIN. EquiPSC clones H, I, and K express all three pluripotency factors while equine fibroblasts (fib) do not. hESC and hiPSC (hOCT4 transgene not silenced) samples verified the specificity of the eqOCT4 primers. eiPSC, equine induced pluripotent stem cells; hESC, human embryonic stem cell; hiPSC, human induced pluripotent stem cell; RT-PCR, reverse-transcription polymerase chain reaction.