摘要
Three methods were adopted in culture spermatogoniums of newly born calf in vitro,such as enzymatic digestion and percoll density gradient centrifugation（MethodⅠ）,tubular fragments culture（MethodⅡ）and tissue culture（MethodⅢ）,and cultural behaviors of cells were observed.The results showed that typical spermatogonium colonies appeard at 144 h of culture by enzymatic digestion-percoll density gradient centrifugation method and tubular fragments culture method,2.5%FBS kept the characteristics of spermatogonium stem cell better than others,produced more mass clones,and FBS of more than 2.5%concentration benefited spermatogonium differentiation and the number of colonies was significantly affected by FBS concentration.After 1 week of culture in method Ⅲ,the diameter of lumens and quantity of sertoli’s cells in tubal wall increased obviously,lumen of seminiferous tubules appeared.Sertoli’s cells kept constant and the number of spermatogoniums decreased obviously after 2 weeks of culture.
Three methods were adopted in culture spermatogoniums of newly born calf in vitro,such as enzymatic digestion and percoll density gradient centrifugation（MethodⅠ）,tubular fragments culture（MethodⅡ）and tissue culture（MethodⅢ）,and cultural behaviors of cells were observed.The results showed that typical spermatogonium colonies appeard at 144 h of culture by enzymatic digestion-percoll density gradient centrifugation method and tubular fragments culture method,2.5%FBS kept the characteristics of spermatogonium stem cell better than others,produced more mass clones,and FBS of more than 2.5%concentration benefited spermatogonium differentiation and the number of colonies was significantly affected by FBS concentration.After 1 week of culture in method Ⅲ,the diameter of lumens and quantity of sertoli’s cells in tubal wall increased obviously,lumen of seminiferous tubules appeared.Sertoli’s cells kept constant and the number of spermatogoniums decreased obviously after 2 weeks of culture.
基金
Supported by Fund of Heilongjiang Acadamy of Agricultural Science
