Isolation, Purification And Culture Of Spermatogonia In Chicken

To ascertain the best group of enzymatic digestion and the best obtained time of spermatogonia, chicken testicular tissues were digested by different enzymatic digestion group and the purity of spermatogonia obtained from four different time periods were also compared. In the condition of purity, the single cell suspension was further treated by using percoll gradient centrifugation and adhesion purification after an enzymatic digestion in order to explore the suitable method of chicken spermatogonia isolation and purity; In the condition of culture, this study compared the different condition of chicken spermatogonia cultured in have-no CEF feed layer and have-no somatic cells, the aim was explore the suitable method and ways for the culture and establish lines in vitro of spermatogonia. The results showed that:1. Using three methods of"1mg/ml colleagenase +0.25% trypsin +0.25% trypsin","1mg/ml colleagenase +1.5mg/ml Hyaluronidase /0.25% trypsin"and"1mg/ml colleagenase +grinding+0.25% trypsin"to digest chicken testicular tissues and found that: the treatment of 1mg/ml colleagenase I combined with 0.25% trypsin was better and the rate of total living cells and the number of living cells in every testis was also higher, which was up to 94.5% and 3.24 105 respectively. When isolate and purify spermatogonia from chicken testicular tissues of different periods and found that: the purity of spermatogonia (61.9%) obtained from 6d-old chicken was markedly higher than of embryonic periods and 13d-old chicken. Result shows that: high rate of living cells and a large number of living cells in every testis and spermatogonia can be...