摘要
利用密度梯度离心从孵化51～56h的鸡胚血液中提纯PGCs，通过形态鉴别和过碘酸－希夫试剂染色方法判定提纯的PGCs的纯度在80％左右。将提取到的PGCs放入到含有10％二甲基亚砜的TCM－199中，在液氮中进行保存。将超低温冷冻保存了1个月、6个月和12个月的PGCs（各3管）分别复苏后，用台盼蓝染色，通过红细胞计数板计数鉴定冷冻保存了不同时间的PGCs的平均存活率分别为91．7％、92．6％和91．9％。结果表明所采用的冷冻保存的方法是可行的。所采用的超低温冷冻保存方法简单易行，为今后以PGCs为目的细胞进行超低温冷冻，而后制作种系嵌合体进行保种的研究提供有利的实践基础。
Primordial germ cells （PGCs） were purified from the blood of chickens embryos hatched for 51 ～ 56 hours by Ficoll density gradient centrifugation. The purity of PGCs could be up to approximately 80% by identification of configuration and periodic acid - schiff （PAS） dyeing. Purified PGCs were suspended in TCM - 199 containing 10% dimethyl sulfoxide and preserved in liquid nitrogen. The average survival rats of frozen - thawed PGCs with different preservation time of 1, and 91.7% 6 and 12 months in liquid nitrogen were 91.7%, 92.7% respectively, by identification of trypan blue staining. The results indicated that the freeze - preservation method of PGCs using in this experiment was simple and feasible. It provided an effective practical groundwork to produce germine chimera for conservation of genetic material by transferring of PGCs preserved in liquid nitrogen.
出处
《江西农业大学学报》
CAS
CSCD
北大核心
2008年第6期981-985,共5页
Acta Agriculturae Universitatis Jiangxiensis
基金
湖南科技大学资助项目(E50477)
关键词
鸡
原生殖细胞
超低温保存
chicken
primordial germ cells （PGCs）
ultra -low temperature storage
