不同冷冻剂对鸡胚PGCs的保护效应  

Effect of different cryoprotectants on the cryopreservation of chicken EPGCs

作　　者：韩威[1] 李碧春[1] 秦洁[1] 朱云芬[1] 陈国宏[1] 

机构地区：[1]扬州大学动物科学与技术学院,江苏扬州225009

出　　处：《中国兽医学报》2007年第3期416-419,共4页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30170678)

摘　　要：发育至第19期和第28期的鸡胚性腺原始生殖细胞(PGCs)分离提纯后,在DMEM中添加冷冻保护剂DMSO(二甲基亚砜)、EG(乙二醇)、蔗糖进行超低温冷冻保存,比较冷冻保护剂在单独或联合使用条件下对PGCs的冷冻保护效果,复苏后台盼蓝染色测定细胞存活率,体外接种培养、传代。结果表明:(1)第19期PGCs冷冻保存中,10% EG+10% FBS+0.1mol/L蔗糖条件下细胞存活率最高为(92.20±2.18)%,且与10% DMSO+10% FBS的存活率差异显著(P<0.05),其余各冷冻保护液间存活率差异不显著;第28期PGCs冷冻保存中,5% DMSO+5% EG+10% FBS+0.1mol/L蔗糖条件下细胞存活率最高为(92.41±2.82)%,但各冷冻保护液间存活率差异均不显著(P>0.05)。(2)解冻后体外培养的PGCs,在饲养层和3种细胞因子(LIF、SCF、bFGF)的共同作用下,可持续增殖,传至第3代的PGCs,PAS染色、AKP染色均呈阳性并保持完整的二倍体核型,仍处于未分化状态。This paper focused on isolated primordial germ cells （PGCs） from gonads at stage 19 and stage 28 by Ficoll density-gradient centrifugation. Then,the PGCs were frozen in five kinds of freezing media under the freezing process. The viability of the frozen-thawed PGCs was measured by the trypan blue exclusion method. The results showed ;@For the stage 19,the viability of the frozen-thawed PGCs was significant difference （P〈0.05） between freezing media I （10% DMSO＋10% FBS） and freezing media IV （10% EG-kl0% FBS＋0.1 mol/L sucrose）,but that of the stage 28 showed no significant difference（P〈0.05） among all the freezing media. ②When cultured on feeder cells in the presence of the three kinds of cytokines,the third passage PGCs still can proliferate and maintain its undifferentiated state.

关 键 词：鸡 原始生殖细胞(PGCs) 冷冻保存 培养传代 

分 类 号：S814[农业科学—畜牧学]