Yan A-Yong, Li Ming, An Xiao-Rong, Hou Jian, Guan Hong, Chen Yong-Fu, Gou Ke-Mian
State Key Laboratories for Agrobiotechnologies, College of Biological Sciences, China Agricultural University, Beijing 100094, China.
Sheng Wu Gong Cheng Xue Bao. 2005 Mar;21(2):305-10.
This paper describes the use of piezo-driven micropipette for intracytoplasmic sperm injection of mice eggs. The head of fresh spermatozoa from KM (Kunming) fertile mice was individually injected into mature oocytes of hybrid mice B6D2F1. Approximately eighty three percent of sperm-injected oocytes 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 sperm 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 from 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 embryos 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. Except 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.
本文描述了使用压电驱动的微量移液器对小鼠卵子进行胞浆内精子注射的情况。将来自KM(昆明)可育小鼠的新鲜精子头部逐个注射到杂交小鼠B6D2F1的成熟卵母细胞中。约83%的注射精子的卵母细胞存活,其中84.0%正常受精(第二极体排出以及雌雄原核形成)。通过精子注射受精的卵子在体外可发育到2细胞期(98%对94.7%)、4细胞期(89.5%对92.1%),与体内受精的胚胎相比无显著差异(P>0.05),但在ICSI组中,进一步体外培养后,体外发育的桑椹胚(63.8%对84.2%)和囊胚(25.7%对68.4%)显著减少(P<0.01)。将120个原核期胚胎移植到7只假孕的KM雌性小鼠体内,23.3%的胚胎(0 - 50%,取决于宿主)发育至足月。除了3只出生后不久被同类相食外,所有幼崽(25只幼仔)都发育成了正常且可育的成年个体。在此我们报道中国首例ICSI后的小鼠后代出生情况。这些研究可能会增进对受精过程以及ICSI作用机制的理解。