[Factors influencing isolation and cloning of chicken primordial germ cells]

Gonadal primordial germ cells (PGCs) were isolated from 4.5-7.5-day-old chicken embryos. Factors influencing isolation and cloning of chicken PGCs were analyzed in this study. The results attained were as follows: Chicken PGCs proliferated fast in presence of 10% FBS. At primary culture, PGCs were co-cultured with the gonadal stromal cells. For subculture the chicken PGCs were plated with mitotically inactive feeder layer cells. Further research found that the PMEF (primary mice embryonic fibroblast) feeder cells were the most suitable for the growth of PGCs. When the cell culture medium was supplemented with LIF, FGF, SCF and IGF-I, the chicken PGCs were in the best condition. The chicken PGCs were easier to clone when isolated from 5.5-day-old embryonic gonads. In addition, the proper digestion time was 40s when PGCs colonies were passaged.