Culture of Chicken Gonadal Primordial Germ Cells (gPGCs) in Chicken Embryonic Fibroblast (CEF) Cells Conditioned Medium and in vivo Migration

The primitive gonads collected from 5.5 days old chicken embryos were dissociated and explanted onto plates pre-coated with 0.1% gelatin. The conditioned media from proliferating and Chicken Embryonic Fibroblast (CEF) cells was supplemented with growth factors and used to support the growth of gPGCs. The result showed that the conditioned media could promote the growth and colony formation of gPGCs in vitro in particular the conditioned by CEF cells. The gPGC derived colonies maintained in CEF cells-conditioned medium up to 281 days were positively antibodies specific to anti-SSEA-1, SSEA-4 integrin alpha 6 and integrin beta 1 and stained by periodic acid Schiff reaction. Their capacities of migration via vascular system and taking up residence in the primary gonadal ridge were further demonstrated by transferring to the dorsal aorta of stage 17 recipient embryos. These results suggested that the culture system is able to maintain chicken gPGCs for long-term in vitro culture without losing their capacity to express pluripotent markers and to integrate into the gonads.