Saadeldin Islam M, Jang Goo
Department of Animal Production, College of Food and Agricultural Sciences, King Saud University, 11451 Riyadh, Saudi Arabia.
Department of Physiology, Faculty of Veterinary Medicine, Zagazig University, 44519 Zagazig, Egypt.
Int J Vet Sci Med. 2018 Feb 15;6(Suppl):S78-S80. doi: 10.1016/j.ijvsm.2018.01.003. eCollection 2018.
Single embryo culture is essential for culturing embryos derived from few oocytes obtained from elite cows through ultrasonography guidance. Bovine fertilization (IVF) and individual embryo culture is a challenge as it generally leads to impaired embryo development. In this study, we explored the embryonic development and the sex ratio of IVF-derived bovine embryo cultured individually in chemically defined two-step culture medium. Total 63 cumulus-oocyte complexes were collected, matured, fertilized and the resultant fertilized oocytes were randomly cultured individually (4 trials, 15-16 oocytes each) in microdrops of 5 µL of a chemically defined two-step culture medium. Blastocysts were counted in every trial (n = 32, 50.79%) and all of them were used for both genomic DNA and total RNA extraction, cDNA synthesis and PCR using specific primers for , , and genes. Results showed significant difference in expression of (positive expression in 11 blastocysts) and (positive expression in 21 blastocysts) mRNAs, < .05. This result supports the hypothesis of sexual dimorphism among the pre-implantation produced embryos and provides an efficient medium for single bovine embryos production.
单胚胎培养对于培养通过超声引导从优质奶牛获取的少量卵母细胞所得到的胚胎至关重要。牛的体外受精(IVF)和单个胚胎培养是一项挑战,因为这通常会导致胚胎发育受损。在本研究中,我们探索了在化学成分明确的两步培养基中单独培养的IVF衍生牛胚胎的胚胎发育和性别比例。总共收集了63个卵丘 - 卵母细胞复合体,使其成熟、受精,然后将所得的受精卵在5微升化学成分明确的两步培养基的微滴中单独随机培养(4次试验,每次15 - 16个卵母细胞)。在每次试验中对囊胚进行计数(n = 32,50.79%),并且所有囊胚都用于基因组DNA和总RNA提取、cDNA合成以及使用针对 、 、 和 基因的特异性引物进行PCR。结果显示 和 mRNA的表达存在显著差异(分别在11个和21个囊胚中呈阳性表达), < 0.05。这一结果支持了植入前产生的胚胎中存在性别二态性的假设,并为单个牛胚胎生产提供了一种有效的培养基。