Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, ON, Canada, N1G 2W1.
Hum Reprod. 2010 Feb;25(2):334-44. doi: 10.1093/humrep/dep394. Epub 2009 Nov 17.
Early embryo development (EED) forms the basis of assisted reproductive technologies (ARTs), which are used to treat human infertility and to propagate other mammalian species. Thyroid hormones (THs) play an important role in the post-implantation development of the embryo in mammals; however, the effects of THs on pre-attachment embryos are not known. Currently utilized in-vitro embryo production media are devoid of THs and hence our main objective was to examine whether THs affected EED in a bovine model.
To determine if THs are present at the site of fertilization and EED in cattle, we evaluated the presence of the hormones in oviductal and uterine horn tissues. To assess the outcome of free TH supplementation (50 ng/ml of each hormone: triiodothyronine-T3 and thyroxin-T4), embryos were followed through standard and TH-supplemented in-vitro procedures, and evaluated for the cleavage rates, blastocyst formation rate and hatching rates. Embryo quality was assessed using TUNEL assay and post-cryopreservation survival was also evaluated.
Although TH levels in in-vitro culture media were found to be approximately 60% of the administered doses, the TH-treated embryos exhibited significant increases in blastocyst formation and hatching rates (P < 0.05). Embryo quality was significantly improved in the treated groups as demonstrated by greater total cell counts and reduced proportions of apoptotic cells (P < 0.05). Finally, TH supplementation was associated with improved post-cryopreservation viability, defined by blastocyst re-expansion and hatching rates after frozen embryos had been thawed and cultured (P < 0.05).
These findings not only provide a way of optimizing ART efficiency, but also further our understanding of how THs influence embryonic development in mammals.
早期胚胎发育(EED)是辅助生殖技术(ARTs)的基础,这些技术用于治疗人类不育症和繁殖其他哺乳动物物种。甲状腺激素(THs)在哺乳动物胚胎的着床后发育中起着重要作用;然而,THs 对附植前胚胎的影响尚不清楚。目前用于体外胚胎生产的培养基中缺乏 THs,因此我们的主要目标是研究 THs 是否会影响牛模型中的 EED。
为了确定 THs 是否存在于受精和牛的 EED 部位,我们评估了激素在输卵管和子宫角组织中的存在。为了评估游离 TH 补充(每种激素 50ng/ml:三碘甲状腺原氨酸-T3 和甲状腺素-T4)的结果,胚胎通过标准和 TH 补充的体外程序进行跟踪,并评估其分裂率、囊胚形成率和孵化率。使用 TUNEL 测定评估胚胎质量,还评估了冷冻保存后的存活情况。
尽管体外培养基中的 TH 水平被发现约为给予剂量的 60%,但 TH 处理的胚胎显示出囊胚形成和孵化率的显著增加(P < 0.05)。处理组的胚胎质量显著提高,表现为总细胞计数增加和凋亡细胞比例降低(P < 0.05)。最后,TH 补充与冷冻保存后活力的改善相关,定义为冷冻胚胎解冻和培养后囊胚的再扩张和孵化率(P < 0.05)。
这些发现不仅提供了优化 ART 效率的方法,而且进一步加深了我们对 THs 如何影响哺乳动物胚胎发育的理解。