第28期鸡胚原始生殖细胞的冷冻保存与体外培养研究  

Cryopreservation and culture of chicken primordial germ cells at stage 28

作　　者：肖小珺[1] 蔡琳琳[1] 秦洁[1] 李碧春[1] 

机构地区：[1]扬州大学动物科学与技术学院

出　　处：《西北农林科技大学学报（自然科学版）》2005年第2期39-42,48,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家自然科学基金资助项目(30170678)

摘　　要：　利用Ficoll密度梯度离心,提取鸡胚第28期(孵化132h)性腺中的原始生殖细胞(PGCs),用不同的冷冻保护液,对PGCs采用分离后直接冷冻保存和体外培养24h后冷冻保存2种方法,以筛选合适的冷冻保护体系。结果发现,分离提纯后直接进行冷冻保存的PGCs,存活率最高为(87.07±1.29)%;体外培养24h后进行冷冻的PGCs,存活率最高为(44.08±1.19)%。对复苏后的PGCs进行体外培养结果发现,分离提纯后直接进行冷冻的PGCs,在体外培养过程中具有形成细胞克隆的能力,且可传代和进行体外分化;而体外培养24h后进行冷冻的PGCs,复苏后在体外培养过程中不能形成细胞克隆,且在体外培养40h左右死亡。In order to investigate the possibility of cryopreservation and differentiation of the chicken primordial germ cells (PGCs) at stage 28,the PGCs from the gonada by Ficoll density-gradient centrifugation were isolated,and cryopreserved right after isolation or after being cultured 24 h in vitro in different preservatives.The vitality of the frozen-thawed PGCs was determined by Trypan blue exclusion method.The result showed:the vitality was the highest (87.07±1.29)% when the PGCs cryopreserved right after isolation.The thawed PGCs could form colony when cultured in vitro.The vitality of the PGCs reached highest (44.08±1.19)% when it was cultured 24 h in vitro before cryopreservation,and the PGCs could not form colony and could only survive about 40 h when culture in vitro.

关 键 词：鸡 原始生殖细胞 冷冻保存 体外培养 

分 类 号：Q813.7[生物学—生物工程]