Size-specific follicle selection improves mouse oocyte reproductive outcomes.
Size-specific follicle selection improves mouse oocyte reproductive outcomes.
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DOI：
10.1530/rep-15-0175
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发表时间：
2015-09
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影响因子：
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通讯作者：
Woodruff TK
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文献类型：
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作者：
Xiao S;Duncan FE;Bai L;Nguyen CT;Shea LD;Woodruff TK
包埋式体外卵泡生长技术(EIVFG)为患有癌症或其他生殖健康疾病的年轻妇女和女孩提供了一种额外的生育保护选择，具有巨大的潜力。目前，卵泡被培养一段确定的时间，并作为一个队列进行分析。然而，卵泡的生长不是同步的，培养时间不足或过多都会导致配子质量下降。我们的目标是确定基于大小而不是绝对培养时间来选择卵泡是否能更好地预测卵泡的成熟度和卵母细胞质量。分离小鼠多层次级卵泡，用0.25%海藻酸盐包裹。分别培养卵泡不同的时间段或特定的卵泡直径范围，在这一点上分析几个生殖终点。培养特定天数的卵泡，第6天卵母细胞成熟后中期II(MII)百分率最高(85%)。然而，如果将卵泡培养到末端直径300-350μm，而不考虑培养的绝对时间，93%的卵母细胞达到MII。从直径30 0~35 0μm的卵泡中成熟的MII卵母细胞中，90%以上的卵母细胞具有正常的纺锤体形态和染色体排列，85%的卵母细胞体外受精后出现2个原核，81%发育到2-细胞胚胎阶段，38%发育到囊胚期，均显著高于其他卵泡大小组。我们的研究表明，在eIVFG过程中，特定大小的卵泡选择可以作为识别高质量卵母细胞和改善生殖结局的非侵入性标记。
Encapsulated in vitro follicle growth (eIVFG) has great potential to provide an additional fertility preservation option for young women and girls with cancer or other reproductive health threatening diseases. Currently, follicles are cultured for a defined period of time and analyzed as a cohort. However, follicle growth is not synchronous, and culturing follicles for insufficient or excessive times can result in compromised gamete quality. Our objective is to determine whether the selection of follicles based on size, rather than absolute culture time, better predict follicle maturity and oocyte quality. Multilayer secondary mouse follicles were isolated and encapsulated in 0.25% alginate. Follicles were cultured individually either for defined time periods or up to specific follicle diameter ranges, at which point several reproductive endpoints were analyzed. The metaphase II (MII) percentage after oocyte maturation on day 6 was the highest (85%) when follicles were cultured for specific days. However, if follicles were cultured to a terminal diameter of 300–350 μm irrespective of absolute time in culture, 93% of the oocytes reached MII. More than 90% of MII oocytes matured from follicles with diameters of 300–350 μm showed normal spindle morphology and chromosome alignment, 85% of oocytes showed 2 pronuclei after in vitro fertilization (IVF), 81% developed into the 2-cell embryo stage, and 38% developed to the blastocyst stage, all significantly higher than the percentages in the other follicle size groups. Our study demonstrates that size-specific follicle selection can be used as a non-invasive marker to identify high quality oocytes and improve reproductive outcomes during eIVFG.