【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。更多还原

【Abstract】 Six kinds of vitrification freezing medias were used to cryopreserve isolated primordial germ cells(PGCs) from gonads at 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+0.5mol/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.更多还原