Pluripotency marker expression. (A): Test of canine epidermal neural crest stem cell (cEPI-NCSC) molecular characterization primer sets. (B): cEPI-NCSC gene expression after ex vivo expansion. Percentage of relative expression by quantitative reverse transcriptase polymerase chain reaction of pluripotency genes; data from four donors. (C): Characterization at the protein level by immunocytochemistry. cEPI-NCSC show immunofluorescence for the pluripotency markers Oct4 (images A, A′), Sox2 (images B, B′), Klf4 (images C, C′), c-Myc (images D, D′), Lin28 (images E, E′), and Nanog (images F, F′). Blue, DAPI nuclear stain. Data are from five donors. Scale bars = 50 μm. Abbreviations: -, water; -RT, minus reverse transcriptase; DAPI, 4′,6-diamidino-2-phenylindole; E, embryo; EPI, canine epidermal neural crest stem cells; HKG, housekeeping gene; L, standard, 100–200 base pairs.