保护剂浓度对第19期鸡胚原始生殖细胞存活率的影响

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 ,并可传代培养

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

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.