Thi Nguyen Nhien, Hirata Maki, Tanihara Fuminori, Hirano Takayuki, Le Quynh Anh, Nii Masahiro, Otoi Takeshige
Faculty of Bioscience and Bioindustry, Tokushima University, Tokushima, Japan.
Tokushima Prefectural Livestock Research Institute, Tokushima, Japan.
Reprod Domest Anim. 2019 May;54(5):750-755. doi: 10.1111/rda.13417. Epub 2019 Mar 5.
The current study was conducted to investigate the effects of 100% foetal bovine serum (FBS) and 100% porcine follicular fluid (pFF) as a storage medium on the developmental competence of porcine zygotes stored at 25°C for 24 hr. Moreover, we evaluated the additive effects of chlorogenic acid (CGA) in the storage medium. When in vitro-produced zygotes were stored at 25°C for 24 hr in tubes containing either tissue culture medium (TCM) 199 supplemented with 1 mg/ml bovine serum albumin (BSA), 100% of FBS or 100% of pFF, the rate of blastocyst formation was significantly higher in 100% of FBS than in BSA-containing TCM 199. When the effects of CGA supplementation in 100% of FBS on the development of zygotes stored at 25°C for 24 hr was evaluated, more zygotes stored with 50 µM CGA developed to blastocysts compared with the other concentrations of CGA. When the formation date and quality of blastocysts derived from zygotes stored in 100% of FBS supplemented with 50 µM CGA were investigated, the highest ratio of blastocysts formation in the storage group appeared 1 day later than in the non-stored control group. However, a higher proportion of blastocysts with apoptotic nuclei was observed in the stored group as compared to the non-stored group. In conclusion, 100% of FBS is available for a short storage medium of porcine zygotes. The supplementation of 50 µM CGA into the storage medium improves the rates of blastocyst formation of zygotes after storage, but the quality of embryos from the stored zygotes remains to be improved.
本研究旨在探讨100%胎牛血清（FBS）和100%猪卵泡液（pFF）作为储存介质对在25°C下储存24小时的猪受精卵发育能力的影响。此外，我们评估了储存介质中绿原酸（CGA）的添加效果。当体外产生的受精卵在含有补充了1mg/ml牛血清白蛋白（BSA）的组织培养基（TCM）199、100% FBS或100% pFF的试管中于25°C下储存24小时时，100% FBS组的囊胚形成率显著高于含BSA的TCM 199组。当评估在100% FBS中添加CGA对在25°C下储存24小时的受精卵发育的影响时，与其他浓度的CGA相比，添加50µM CGA储存的受精卵发育成囊胚的更多。当研究在补充了50µM CGA的100% FBS中储存的受精卵来源的囊胚形成日期和质量时，储存组囊胚形成的最高比例比未储存的对照组晚1天出现。然而，与未储存组相比，储存组中观察到具有凋亡细胞核的囊胚比例更高。总之，100% FBS可作为猪受精卵的短期储存介质。在储存介质中添加50µM CGA可提高储存后受精卵的囊胚形成率，但储存受精卵来源的胚胎质量仍有待提高。
