Mammalian species utilize an inductive mechanism of germ cell specification, diverting the fate of some of somatic cells toward pluripotency and germ-cell totipotency. It is not known if avian species utilize a similar mechanism nor if, analogous to mammalian primordial germ cells (PGCs), pluripotency genes are continuously upregulated in migrating and genital ridge-colonizing avian PGCs. Thus, this study was conducted to quantify and to analyze the expression profile of pluripotency genes at different stages of chicken PGCs development at Hamburger and Hamilton (HH) stage 14, when the majority of PGCs have entered into the bloodstream; at HH stage 18, when PGCs have resided for 8-12 hr in the bloodstream; and at HH stage 28, when the majority of PGCs are found in the genital ridge. The transcription for Oct4, Sox2, and Nanog continuously decreased from HH stage 14 to HH stage 28. In addition, equal amounts of total RNA could be isolated from chicken PGCs at each stage of development, indicating that the observed drop of transcription of pluripotency genes is not a consequence of transcriptional repression in general. Decreased expression for all three proteins was also observed at HH stage 28. Furthermore, in comparison to blood PGCs, those residing in the gonad have lost their full capacity for colony formation. Our results indicate that, in contrast to mammalian PGCs, chicken PGCs continuously downregulate the expression of pluripotency genes and show a progressive loss of pluripotency-associated features during different stages of germ-line migration.