摘要
以piezo操作系统为技术支撑 ,在掌握小鼠卵母细胞胞浆内精子注射技术 (ICSI)的基础上 ,进行了ICSI技术生产试管小鼠的尝试。来自成年昆明 (KM)小鼠附睾尾的新鲜精子 ,剪切去尾后 ,直接将精子头注射到B6D2F1小鼠卵母细胞质中 ,注射后 1h ,83 .3%的卵母细胞存活。6h时 ,84 . 0 %的成活卵子成功受精 ,形成原核 ,排出PB2 体外培养的ICSI胚胎 ,卵裂率 (98%vs 94 . 7% )和 4_细胞期胚胎比率 (89 5 %vs 92 . 1% )均与培养的体内受精卵没有差异 (P >0 . 0 5 ) ;但是 ,桑椹胚(6 3 8%vs84 . 2 % )和囊胚发育率 (2 5 .7%vs 6 8. 4 % )极显著地 (P <0 . 0 1)低于对照组。12 0枚原核期胚胎移植给 7只假孕受体后 ,4只受孕小鼠共产出 2 8只ICSI小鼠 (2 3. 3% )。健康成年的 2 5只ICSI小鼠都没有明显的生理和行为异常。随机选择其中的 2 0只小鼠 ,分别进行ICSI小鼠间、ICSI与KM小鼠间共 12组的交配 ,结果所有雌鼠妊娠产仔。在成功建立小鼠ICSI技术的基础上 ,成功获得了我国的首例ICSI小鼠 ,并且证明这些ICSI小鼠都具有正常的繁殖后代的能力。
This paper describes the use of piezo-driven micropipe tte for intracytoplasmic sperm injection of mice eggs. The head of fresh spermat ozoa from KM(Kunming)fertile mice was individually injected into mature oocyte s of hybrid mice B6D2F1. Approximately eighty three percent of sperm-injected o o cytes survived, and 84.0 % of them fertilized normally (extrusion of the second polar body and formation of male and female pronuclei). The eggs fertilized by s perm injection could develop in vitro to 2-cell (98 % vs 94.7 %), 4-cell (89.5% vs 92.1 %) stages, no significantly (P>0.05) different fro m embryos fertilized in vivo but there were significantly (P<0.01) few morulae (63.8% vs 84.2 %) and blastocysts (25.7% vs 68.4%) developed in vitro after further culture in vitro in the group of ICSI. When 120 embr yos at the pronuclear stage were transferred to seven pseudopregnant KM female, 23.3% of the embryos (0～50%, depending on the host) reached the full term. Exc ep t for three that were cannibalized soon after birth, all of the young (25 pups) developed into normal and fertile adult. Here we report the first birth of mouse offspring following ICSI in China. These studies may increase understanding of the fertilization process and of how ICSI works.
出处
《生物工程学报》
CAS
CSCD
北大核心
2005年第2期305-310,共6页
Chinese Journal of Biotechnology
基金
国家自然科学基金 (No .3 0 2 70 64 7
3 0 2 70 95 6)
北京市自然科学基金 (No .5 0 12 0 0 7)
国家"863"高技术研究发展计划 (No.2 0 0 2AA2 0 63 11)资助项目~~
关键词
ICSI
囊胚
小鼠
生殖力
mouse, intracytoplasmic sperm injection (ICSI),off spring
