小鼠卵母细胞MII期核(纺锤体)移植后经体外受精正常产仔

In vitro fertilization of murine oocytes with metaphase II apparatus (spindle) transfer results in live births

目的 :为研究和了解细胞核与细胞质的相互作用。方法 :采用 3 %蔗糖预处理显示及去核法进行昆明白小鼠与 C5 7BL / 6小鼠 (黑色 )之间卵母细胞 MII期核 (纺锤体 )的移植、电融合、体外受精 -胚胎移植 ( IVF-ET)。结果 :经 3 %蔗糖预处理卵母细胞的去核率达 1 0 0 %。完成 MII期核移植的重构卵母细胞约 1 7.6%完成电融合。IVF的受精率为82 %。受精卵 2 -细胞胚胎的发育率为 80 %～ 90 % ,8个 1～ 4细胞胚胎移入两只自然怀孕1 d的寄母小鼠 ( Km× Km)输卵管 ,其中 1只孕鼠生下 3只小鼠 ,其中 2只黑色小鼠为MII期核 (纺锤体 )移植生产的小鼠。移植产仔率为 2 5 %。结论 :实验结果证明了 3 %蔗糖预处理显示 MII期核 (纺锤体 )及去核效果可达 1 0 0 % ;电融合的重构卵母细胞可完成正常的 IVF。为进行不同年龄和卵龄间的

To study the interaction between nucleus and cytoplasm. Methods: Operation medium containing 3% sucrose was used to pretreatment murine oocytes and meiotic apparatus transfer(MAT) was carried out from C57BL/6 mouse (black) to enucleated Kunming murine oocytes and followed by electrofusion, in vitro fertilization (IVF) and embryo transfer. Results: The results showed that the enucleation rate by treating oocytes with 3% sucrose was 100%, 17.6% of reconstructed karyoplasm recipient cytoplasm pairs fused sucessfully. When the fused oocytes were exposed to spermatozoa from C57BL/6 mice, 9 out of 11 (82%) were fertilized. Among them, eight embryos at one to four cell stages were transferred into the oviducts of two synchronous pregnant Kunming strain fosters and one delivered two normal C57BL/6 offsprings. Conclusion: This study indicated that MII meiotic apparatus or spindle sustains normal structure and function after micromanipulation and electrofusion. MAT provides a model for further research on the application of this technique to assistant human reproduction.