PSVII-19 Characterization and Migration of Cultured Quail Primordial Germ Cells from Embryonic Blood and Gonad.

摘要

Primordial germ cells (PGCs), the precursors of ova and spermatozoa are ideal tools for transgenic technology. Therefore isolation, characterization and migration of PGCs is required. The quail has been proven to be a good model in numerous fields of research, especially for biotechnology due to short generation time. In this study, initially we attempted to isolate quail PGCs from gonad (at stage 28–30) and blood (at stage 13–15) as two sources of these cells. The isolated PGC were cultured on feeder cells derived from chicken embryonic fibroblast. Then characterization of these cells were evaluated during the culture. In finally for migration assay of these cells into the gonads, cultured PGCs were injected into blood circulate of the recipients quail embryos. The cultured gonadal PGC proliferated about 400-times versus 100-times in blood PGC during the first 40–50 days. The identities of the PGC were confirmed using periodic acid-Schiff (PAS) staining or anti-SSEA1, β-Catenin, β-integrin, and Nanog immunofluorescence staining or reverse transcription-polymerase chain reaction for several factors related to PGC phenotype, , , and . Cultured PGCs migrated toward the embryonic gonads when they were transplanted into the blood circulate of the recipient embryo at stage 13–15 Hamburger–Hamilton (HH). Our results suggest that cultured PGCs retain their ability to migrate and colonize the gonads of recipient embryos.