Comparison of spindle and chromosome configuration in in vitro- and in vivo-matured mouse oocytes after vitrification
Comparison of spindle and chromosome configuration in in vitro- and in vivo-matured mouse oocytes after vitrification
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DOI：
10.1016/j.fertnstert.2007.07.1335
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发表时间：
2008-10-01
影响因子：
6.7
通讯作者：
Chian, Ri-Cheng
中科院分区：
文献类型：
--
作者：
Huang, Jack Y. J.;Chen, Hai Ying;Chian, Ri-Cheng
目的：比较体外和体内成熟卵母细胞玻璃化冷冻后的细胞遗传学变化。设计：采用小鼠模型进行体外实验。单位：大学实验室动物模型研究。动物(S)：CD-1小鼠。干预(S)：卵母细胞的体外成熟和玻璃化冷冻。主要观察指标(S)：温育后存活率、纺锤体和染色体构型分析、孤雌激活卵母细胞非整倍体筛查、DNA断裂程度、体外受精后早期胚胎发育。结果(S)：生发泡期卵母细胞体外成熟后成熟率为80%，玻璃化冷冻后成熟率为80%。体外和体内成熟卵母细胞升温后存活率分别为94.1%和91.8%，差异无统计学意义(P>0.05)。玻璃化冷冻后，大多数体外和体内成熟的卵母细胞保持了正常的减数分裂纺锤体形态和染色体对齐(分别为88.2%和86.9%)，非整倍体发生率(11.5%和9.3%)没有增加。然而，与体内成熟的卵母细胞相比，体外成熟的卵母细胞在玻璃化冷冻后显示出更高的DNA碎片率。玻璃化冷冻后，体外成熟卵母细胞的卵裂率和囊胚率显著低于体内成熟卵母细胞(分别为37.0%比60.0%和5.4%比18.9%)。结论(S)：玻璃化冷冻小鼠体外成熟卵母细胞存活率高，减数分裂纺锤体和染色体排列正常，非整倍体发生率未见增加。体外成熟和玻璃化冷冻的卵母细胞发育能力下降的一个可能原因可能是DNA片段化。
Objective: To compare the cytogenetic changes in in vitro- and in vivo-matured oocytes after vitrification. Design: In vitro experiments using murine model. Setting: Animal model study in university laboratory. Animal(s): CD-1 mice. Intervention(s): In vitro maturation and vitrification of oocytes. Main Outcome Measure(s): Post-warming survival, analysis of spindle and chromosome configurations, aneuploidy screening of parthenogenetically activated oocytes, extent of DNA fragmentation, and early embryonic development after IVF. Result(s): Eighty percent of germinal vesicle-stage oocytes matured after in vitro maturation and were cryopre-served by vitrification (n = 354). There was no significant difference in the post-warming survival of in vitro- and in vivo-matured oocytes (94.1% vs. 91.8%, respectively). The majority of in vitro- and in vivo-matured oocytes maintained normal meiotic spindle morphology and chromosome alignment (88.2% vs. 86.9%, respectively) after vitrification and the incidence of aneuploidy was not increased (11.5% vs. 9.3%). However, in vitro-matured oocytes showed a higher rate of DNA fragmentation after vitrification compared to in vivo-matured oocytes. After vitrification, the cleavage and blastocyst formation rates of in vitro-matured oocytes were significantly lower than those of in vivo-matured oocytes (37.0% vs. 60.0% and 5.4% vs. 18.9%, respectively). Conclusion(s): Vitrification of in vitro-matured mouse oocytes results in high survival rates, normal meiotic spindle and chromosome alignment, and no increased incidence of aneuploidy. A possible cause of the reduced developmental competence of in vitro-matured and vitrified oocytes may be due to DNA fragmentation.