Isolation and Culture of Primordial Germ Cells of Chicken Embryo at 28th Stage

[Objective] The study aimed to optimize the conditions for culture in vitro of primordial germ cells(PGCs).[Method] PGCs were isolated from the genital ridges of chicken embryos at the 28th stage(5.5 d),then they were co-cultured with gonad stroma cells for primary culture.The effect of 2 kinds of culture temperatures and 3 kinds of culture concn.on PGCs primary culture and that of 2 kinds of feeder layer cells on PGCs subculture were compared.PGCs were identified by morphologic observation under inverted microscope and staining of alkaline phosphatase(AKP) and periodic acid-schiff regent(PAS).[Result] PGCs could proliferate,maintain undifferentiated situation,and had stronger viability when the cell culture concn.was 2.5×105/ml and the culture temperature was 37 ℃.The second passage of chicken embryo fibroblast(CEF) as the feeder layer was more suitable for PGCs subculture and got the undifferentiated colony of PGCs proliferation,which afforded the ample PGCs with high viability for the subsequent experiment of PGCs labeling and transplantation.[Conclusion] The culture system with better primary culture and subculture for PGCs was obtained,which laid the foundation for establishment of avian cell line and its transgenic research.