保护剂浓度对第19期鸡胚原始生殖细胞存活率的影响

Cryopreservation of Chicken Primordial Germ Cells at Stage 19 Under Different Preservative Density

采用Ficoll密度梯度离心 ,提取第 19期 (孵化 72h)性腺中的原始生殖细胞 (PGCs) ,用不同浓度的冷冻保护液进行冷冻保存。复苏后的PGCs用台盼蓝染色检测其存活率 ,并进行体外培养。结果发现 :在同一浓度下 ,不同的冷冻保护液之间存在显著 (P <0 0 5 )或极显著 (P <0 0 1)差异。在同一种冷冻保护液下 ,不同的浓度之间存在显著(P <0 0 5 )或极显著 (P <0 0 1)差异 ,具体表现为冷冻保护液浓度越大 ,复苏后细胞的存活率越低。PGCs复苏后于体外培养 ,可增殖形成细胞克隆 。

Isolating the primordial germ cells(PGCs)from gonads at stage 19 by Ficoll density gradient centrifugation.Cryopreservating the PGCs at difference freezing media density.The vitality of the frozen-thawed PGCs was determined by the Trypan blue exclusion method,than the PGCs were cultured in vitro.The result showed:at the same density,there is significant difference(P<0.05) or very significant difference(P<0.01) among the freezing media.At the same freezing medie,there is significant difference(P<0.05)or very significant difference(P<0.01) among the different density,the density more higher,the vitality more lower.When the thawed PGCs culture in vitro,it can propagation,form PGCs colony,subculture.