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双极囊胚

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In the present study, we collected cumulus cells oocyte complex from ovaries of two different strain mice. The cumulusenclosed oocytes were cultured for 6 h in MEM supplemented with growth factor and FSH. The meiotic maturation of these oocytes has progressed to pro-metaphse Ⅰ stage and the condensed chromosomes are visible under DIC microscope, metaphase Ⅰ spindle even can be detected under Polscope. The metaphase Ⅰ spindles of oocytes were exchanged under such microscopes. After electric stimuli, 91. 6% and 91. 6% karyoplasts-cytoplasm pairs were fused respectively. The resulting oocytes were cultured further in MEM and over 80% of oocytes released the first polar body. 79% and 77% of oocytes formed two pronuclei after in vitro fertilization and the embryos were cultured in KSOM supplemented with amino acids. Over 60% of embryos developed to blastocyst stage.

在本研究中我们在取得两种不同品系小鼠的卵丘卵母细胞复合体后,先将卵丘卵母细胞复合体置于含有多种生长因子和激素的MEM培养液中培养6小时,此时卵母细胞已进入第一次减数分裂的前中期,并且在DIC倒置显微镜下可以看到浓缩的染色体,用Polscope可以发现明显的纺锤体,借助这种显微镜通过显微操作将两种不同品系小鼠来源的卵母细胞的MI纺锤体进行互换,经过三次直流电脉冲作用后,分别有91.6%的胞质—MI核质体对融合,经过进一步的培养后,超过80%的重组卵母细胞排出第一极体,体外受精后分别有79%和77%的重组卵形成双原核,受精后的胚胎在KSOM胚胎培养液中体外培养4天后,超过60%的胚胎发育至囊胚。

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