鸡胚原始生殖细胞(PGCs)玻璃化冷冻的研究  

Effect of Vitrification on Chicken PGCs

作　　者：韩威[1] 李碧春[1] 陈国宏[1] 秦洁[1] 朱云芬[1] 周冠月[1] 张学余[2] 

机构地区：[1]扬州大学动物科学与技术学院,扬州225009 [2]中国农业科学院家禽研究所,扬州225003

出　　处：《畜牧兽医学报》2007年第3期301-306,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金资助(30170678)

摘　　要：对发育至第19期和第28期鸡胚性腺原始生殖细胞(Primordial germ cells,PGCs),用6种玻璃化冷冻液I:10%DMSO+10%EG+10%PVP,II:20%EG+10%PVP,III:20%DMSO+10%PVP,IV:10%DMSO+10%EG+0.5 mol/L Surcose,V:20%EG+0.5 mol/L Surcose,VI:20%DMSO+0.5 mol/L Surcose进行冷冻保存。结果:第19期鸡胚PGCs玻璃化冷冻复苏后存活率,在冷冻液II与IV之间差异不显著(P>0.05),V与VI之间差异显著(P<0.05),其余各冷冻液之间差异均极显著(P<0.01)。第28期鸡胚PGCs玻璃化冷冻复苏后存活率,在冷冻液IV与V之间差异显著(P<0.05),III与IV、V之间差异不显著(P>0.05),II与III、IV之间差异不显著(P>0.05),其余各玻璃化冷冻液之间差异均极显著(P<0.01)。复苏后接种培养传至第2代的鸡胚PGCs细胞,PAS染色、AKP染色呈阳性并保持完整的二倍体核型。Six kinds of vitrification germ cells （PGCs） from gonads at freezing medias were used to cryopreserve isolated primordial stage 19 and stage 28 by ficoll density-gradient centrifugation. The six kinds of vitrification freezing medias were following： Ⅰ ： 10%DMSO＋10%EG＋10% PVP, Ⅱ ： 20% EG ＋ 10% PVP, Ⅲ ： 20% DMSO ＋ 10% PVP, Ⅳ： 10% DMSO ＋ 10% EG ＋ 0. 5 mol/L surcose, Ⅴ ：20%EG＋0.5mol/L surcose, Ⅵ ：20%DMSO＋0.5 mol/L surcose. The viabilities of PGCs were performed after thawing by way of staining with trypan blue. The results showed： For the viability of the frozen-thawed PGCs at stage 19, it showed significant difference （P〈0.05）between the viability under freezing media Ⅵ and the viability under freezing media Ⅴ , and showed insignificant difference （P〉0.05）between the viability under freezing media Ⅱ and the viability under freezing media Ⅳ. But it showed very significant difference（P〈0.01） among the viabilities under the other freezing medias. For the viability of the frozen-thawed PGCs at stage 28, it showed significant difference（P（0.05）between the viability under freezing media Ⅳ and the viability under freezing media Ⅴ , and showed insignificant difference （P〉0. 05） among the viability under freezing media Ⅲ, Ⅳand Ⅴ , and so were the viabilities under freezing media Ⅱ , Ⅲ and Ⅳ （P〈0.05）. But it showed very significant difference（P〈0.01）among the viabilities under the other freezing medias. The second passage PGCs cultured after thawing were positive to PAS staining and AKP staining and still kept intact karyotypes.

关 键 词：鸡 原始生殖细胞 玻璃化冷冻 培养传代 

分 类 号：S831.3[农业科学—畜牧学]