| 摘要: |
| 利用Ficoll密度梯度离心，提取鸡胚第28期(孵化132 h)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24 h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24 h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24 h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40 h左右死亡。 |
| 关键词: 鸡 原始生殖细胞 冷冻保存 体外培养 |
| DOI： |
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| 基金项目:国家自然科学基金资助项目(30170678) |
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| Cryopreservation and culture of chicken primordial germ cells at stage 28 |
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| Abstract: |
| 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. |
| Key words: chicken primordial germ cells cryopreservation culture in vitro |
