Abstract:

A method was described for isolation and culture of pig ear skin fibroblast cells defined for porcine somatic nuclear transfe. The experiment was carried out with pig ear skin fibroblasts for primary culture, routine subculture, freezing, and recovery procedure by tissue-pieced method. Those in vitro cultured cells were analyzed by morphology, growth trends and adherent ability. The results showed that pig ear skin fibroblasts was successfully cultured in vitro efficiently, which was with high growth capacity and short passage-period. After freezing and resuscitation, the fibroblasts could also well attach and proliferate, and could be provided as the appropriate donor cells for related nuclear transfer work. The tissue-pieced method was simple and effective access to the pig ear fibroblasts, the cells could be obtained in vitro for at least 4 generations could be used for donor cells for nuclear transfer embryos.