Laboratory of Animal Reproduction, Graduate School of Veterinary Sciences, Azabu University, Sagamihara, Kanagawa, Japan.
School of Veterinary Medicine, Azabu University, Sagamihara, Kanagawa, Japan.
PLoS One. 2021 Mar 11;16(3):e0248050. doi: 10.1371/journal.pone.0248050. eCollection 2021.
The vitrification of immature germinal vesicle (GV) oocytes is an important way to preserve genetic resources and female fertility. However, it is well known that cryopreserved GV oocytes have very poor developmental ability and that further improvement in this technique is needed. We previously reported the successful vitrification of matured mouse oocytes with enclosed cumulus cells using the calcium-free vitrification solution supplemented with ethylene glycol (EG) by the minimal volume cooling (MVC) method. In this study, we investigated whether our method is applicable to the vitrification of mouse oocytes at the GV stage (GV oocytes). Following maturation and fertilization in vitro, vitrified GV oocytes showed high survival (94.3 ± 2.0%) and maturation (94.3 ± 2.1%) rates. Although the fertilization and blastocyst rates of vitrified oocytes (fertilization: 46.6 ± 4.9% and blastocyst: 46.6 ± 3.0%) were significantly lower than those of fresh oocytes (fertilization: 73.0 ± 7.1% and blastocyst: 71.6 ± 8.0%) (P < 0.01), there were no differences in the ability to develop to term between fresh oocytes (50.0 ± 8.4%) and vitrified oocytes (37.5 ± 4.6%) (P > 0.05). In conclusion, we here show, for the first time, the efficient production of live mice derived from vitrified GV oocytes.
玻璃化法是保存遗传资源和女性生育力的一种重要方法,其中不成熟的生发泡期(GV)卵母细胞的玻璃化尤为重要。然而,众所周知,冷冻保存的 GV 卵母细胞的发育能力非常差,需要进一步改进该技术。我们之前报道了使用无钙玻璃化溶液和乙二醇(EG)通过最小体积冷却(MVC)法成功玻璃化包被卵丘细胞的成熟小鼠卵母细胞。在这项研究中,我们研究了我们的方法是否适用于 GV 期(GV 卵母细胞)小鼠卵母细胞的玻璃化。体外成熟和受精后,冷冻 GV 卵母细胞显示出高存活率(94.3±2.0%)和成熟率(94.3±2.1%)。尽管冷冻卵母细胞的受精率和囊胚率(受精:46.6±4.9%和囊胚:46.6±3.0%)显著低于新鲜卵母细胞(受精:73.0±7.1%和囊胚:71.6±8.0%)(P<0.01),但新鲜卵母细胞(50.0±8.4%)和冷冻卵母细胞(37.5±4.6%)发育至足月的能力没有差异(P>0.05)。总之,我们首次证明了玻璃化 GV 卵母细胞可以高效生产活的小鼠。