In vitro maturation of cumulus-oocyte complexes for efficient isolation of oocytes from outbred deer mice.
In vitro maturation of cumulus-oocyte complexes for efficient isolation of oocytes from outbred deer mice.
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DOI：
10.1371/journal.pone.0056158
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
He X
中科院分区：
文献类型：
--
作者：
Choi JK;He X
近亲繁殖的鹿鼠（和人类一样）已经成为研究人类行为和生物学（包括生殖系统）的有用动物模型。使用该物种的主要挑战之一是，根据迄今为止的文献，通过超排卵分离卵母细胞的产量很低，到目前为止每只动物可以获得不到5个卵母细胞。本研究的目的是通过体外成熟（IVM）累积-卵母细胞复合物（COCs）来提高远交种鹿小鼠的卵母细胞分离率，使其接近大多数实验室小鼠的产率。用超排卵法和体外受精法分离卵母细胞。对于后者，通过对卵巢表面和内皮质层的窦卵泡进行卵泡穿刺获得COCs。然后在优化的条件下体外培养COCs中的未成熟卵母细胞，获得中期II （MII）卵母细胞。通过体外受精（IVF）和胚胎发育检测体外受精和超排卵卵母细胞的质量。每只动物只能通过超排卵分离不到5个卵母细胞。然而，本研究通过IVM（16.0±2.5）和超排卵（4.3±1.3），每只动物成功获得20.3±2.9个卵母细胞。此外，IVF和胚胎发育研究表明，IVM的卵母细胞质量甚至比超排卵的卵母细胞更好，后者通常不会发育到2细胞以上，而前者有9%的卵母细胞发育到4细胞。我们成功地建立了用IVM高产量分离鹿小鼠卵母细胞的方法。此外，这是第一次成功地在体外培养出超过2细胞阶段的体外受精鹿小鼠卵母细胞。因此，本研究对利用鹿鼠进行生殖生物学研究具有重要意义。
The outbred (as with humans) deer mice have been a useful animal model of research on human behavior and biology including that of the reproductive system. One of the major challenges in using this species is that the yield of oocyte isolation via superovulation is dismal according to the literature to date less than ∼5 oocytes per animal can be obtained so far. The goal of this study is to improve the yield of oocyte isolation from outbred deer mice close to that of most laboratory mice by in vitro maturation (IVM) of cumulus-oocyte complexes (COCs). Oocytes were isolated by both superovulation and IVM. For the latter, COCs were obtained by follicular puncture of antral follicles in both the surface and inner cortical layers of ovaries. Immature oocytes in the COCs were then cultured in vitro under optimized conditions to obtain metaphase II (MII) oocytes. Quality of the oocytes from IVM and superovulation was tested by in vitro fertilization (IVF) and embryo development. Less than ∼5 oocytes per animal could be isolated by superovulation only. However, we successfully obtained 20.3±2.9 oocytes per animal by IVM (16.0±2.5) and superovulation (4.3±1.3) in this study. Moreover, IVF and embryo development studies suggest that IVM oocytes have even better quality than that from superovulation The latter never developed to beyond 2-cell stage as usual while 9% of the former developed to 4-cells. We have successfully established the protocol for isolating oocytes from deer mice with high yield by IVM. Moreover, this is the first ever success to develop in vitro fertilized deer mice oocytes beyond the 2-cell stage in vitro. Therefore, this study is of significance to the use of deer mice for reproductive biology research.
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