第十九期鸡胚原始生殖细胞的冷冻保存  

Preservation of chicken primordial germ cells of stage 19 in liquid nitrogen

作　　者：肖小珺[1] 蔡琳琳[2] 秦洁[2] 李碧春[2] 

机构地区：[1]湖州师范学院生命科学院,浙江湖州313004 [2]扬州大学动物科学与技术学院,江苏扬州225009

出　　处：《黑龙江畜牧兽医》2005年第2期7-9,共3页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金(30170678)

摘　　要：采用Ficoll密度梯度离心,提取第 19期(孵化 72h)性腺中的PGCs,对其应用不同的冷冻保护液和不同的平衡方法进行冷冻保存,并于复苏后进行体外培养。复苏后的PGCS用台盼蓝染色检测其存活率,结果发现:从第 19期性腺中获取的PGCs在同一种冷冻保护液下,采用不同的平衡方法进行冷冻,对PGCs的存活率有显著影响(P<0.05)或极显著影响(P<0.01);平衡方法相同,在不同冷冻保护液之间存在显著(P<0.05)或极显著 (P<0.01)差异。PGCs经体外培养 24h后再进行冷冻保存,复苏后其存活率、体外培养存活时间均极显著(P<0.01)短于分离后直接冷冻的PGCs。Isolate primordial germ cells(PGCs) from gonads of stage 19 by Ficoll dencity gradient centrifugation.Cryopreservation the PGCs at difference freezing media and equilibrium method.The viability of the frozenthawed PGCs was determined by the Trypan blue exclusion method,PGCs were cultured in vitro in DMEM medium containing 10%newborn calf serum.The result showed:preserved the PGCs at same freezing media,but at difference equilibrium method,the vitality of the PGCs isolated from gonads of stage 19 or stage 28 showed significant difference or very significant difference;Preserved the PGCs at same equilibrium method,but at difference freezing media,the vitality of PGCs showed significant difference.When the PGCs cryopreserved after 24 h culture in vitro,the survival time showed high significant difference compared to the PGCs cryopreserved directly after isolation.

关 键 词：复苏后 冷冻保护液 冷冻保存 体外培养 存活率 性腺 原始生殖细胞 鸡胚 孵化 平衡方法 

分 类 号：S814[农业科学—畜牧学] R321-33[农业科学—畜牧兽医]