小鼠卵丘细胞核移植到体内成熟卵浆构成的重构卵的发育

[目的]观察小鼠卵丘细胞核移植到体内成熟卵浆构成的重构卵的发育.[方法]昆明小鼠超排后获取体内成熟卵母细胞及卵丘细胞.成熟卵子去核后用将卵丘细胞核与精子直接注入,观察其受精和胚胎发育情况.[结果]卵子去核存活率为75.9%(362/477),注核存活率为39.8%(143/359),D1存活率为38.4%(138/359).39.9%(57/143)重构卵子成功排出一个极体并形成两原核,其卵裂率为80.7%(46/57).大部分重构胚发育到2～3细胞停滞,其中2个分裂至4细胞,无囊胚形成.[结论]将小鼠卵丘细胞核、精子同时移植到小鼠的体内成熟卵浆中,可以诱导极体排出,形成2原核 1极体的重构胚胎.重构胚胎大部分发育到2～3细胞即发生停滞,最多到达4细胞期,发育潜能差.

Development of Reconstructed Mouse Oocyte from In Vivo Matured Ooplasm Transferred with Cumulus Cell Nucleus

To observe the fertilization and development of reconstructed mouse oocyte derived from in vivo matured ooplasm transfered with mouse cumulus cell nucleus. In vivo matured MII oocytes and cumulus cells were collected from stimulated Kunming mice. The nuclei of cumulus cell and spermatozoa were injected directly into the enucleated MII ooplasm. The fertilization and development of reconstructed oocytes were observed. The survival rate after enucleation was 75.9% (362/477); the injection survival rate was 39.8% (143/359); D1 survival rate was 38.4% (138/359). One polar body was extruded and 2 pronuclear formed in 39.9% (57/143) reconstructed oocytes. The cleavage rate was 80.7% (46/57). Most of reconstructed embryos blocked at 2-3 cell stage, only 2 embryos developed into 4-cell, no blast cyst formed. [Conclusion] Mouse reconstructed embryos with 1 polar body and 2 pronuclei can be formed after transferring cumulus cell nucleus and spermatozoa into enucleated MII oocyte. The developmental potential of reconstructed embryo was poor as most of them were blocked at 2-3cell stage and only few could reach 4-cell stage.