2003•Europe PMC (PubMed Central)Requires access
A method for isolation of primordial germ cells in different stages of the chicken embryo
Bichun Li, Jie Qin, Xiaojun Xiao, Guohong Chen, Xinsheng Wu, Shenglong Wu
Open publisher page 1 citations
Abstract
We compared both Ficoll densitYgradient centrifugation and EDT A trypsin collection methods for isolating primordial germ cells(PGCs) from chicken embryos at stage 14, 19 and 28, cultured the PGCs in TCM-199 medium containing 10% FCS without any growth factor under 38.5℃, 5% CO_(2) and saturation moisture to compare the survival time in vitro. The results showed that the survival duration of the PGCs at stage 1 4 was 72h, stagel9 was 88h, and stage 28 was 80h. The number of PGCs, and the viable PGCs with the EDTA trypsin treatment were more(2.71×10~(4), 65.12%)than that of Ficoll density-gradient centrifugation(1.78×10~(4), 45.24%)at stages 19 and 28. Comparison of results of culture of PGCs in vitro at those three development stages indicated that a large amount of PGCs had aggregated in the genital ridge at stage 19 made it convenient for isolation. This stage was suitable for isolation of PGCs from early chicken embryos using EDTA trypsin treatment.