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Similar patterns of embryo development in canine oocytes cultured in vitro at oxygen tensions of 5 and 20%

机译:在5和20%的氧气张力下体外培养的犬卵母细胞中胚胎发育的相似模式

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The influence of two oxygen tensions (5 and 20%) on fertilization, cleavage, development, and morphological quality of canine embryos produced in vitro was investigated. To assess embryo production, canine oocytes (N = 608) were matured in vitro at 20% oxygen tension in TCM 199 supplemented with glucose (11 mM) and 0.1% polyvinyl alcohol. Oocytes and sperm were coincubated at 37 degrees C for 24 hours in serum-free medium. Subsequently, presumptive zygotes were cultured in vitro in synthetic oviductal fluid in either 5% CO2 in air (20% oxygen; N = 298) or 5% O-2, 5% CO2 and 90% N-2 (5% oxygen; N = 310) for 7 days. Regardless of the oxygen concentration (5 vs. 20%), rates of pronucleus formation, cleavage, and embryo development up to the eight-cell stage did not differ (46/310 [14.8%] vs. 35/298 [11.7%], respectively; P > 0.05). Moreover, similar proportions of embryos developed in 20 and 5% oxygen tensions (18/298 vs. 27/310). The oocyte nuclear maturation (metaphase II), in terms of decondensed sperm heads, pronucleus formation, cleavage, and embryo development, was similar (P = 0.299) between the atmospheric (20% O-2; 12.4% [37/298]) and reduced oxygen tensions (5% O-2; 15.8% [49/310]) at all steps of the in vitro culture (IVC) after in vitro fertilization (IVF). To our knowledge, this was the first study that demonstrated that canine embryos can be produced using a low-oxygen in vitro culture system. Both oxygen tensions (5 and 20%) resulted in similar embryonic development and therefore were feasible for IVC of canine oocytes. (C) 2013 Elsevier Inc. All rights reserved.
机译:研究了两种氧气张力(5%和20%)对体外产生的犬胚的受精,分裂,发育和形态质量的影响。为了评估胚胎的产生,犬卵母细胞(N = 608)在补充了葡萄糖(11 mM)和0.1%聚乙烯醇的TCM 199中于20%氧张力下体外成熟。将卵母细胞和精子在无血清培养基中于37度共孵育24小时。随后,假定合子在人工输卵管液中于空气中的5%CO2(20%氧气; N = 298)或5%O-2、5%CO2和90%N-2(5%氧气; N)中进行体外培养。 = 310),持续7天。无论氧气浓度如何(5%vs. 20%),直到八细胞阶段的前核形成,分裂和胚胎发育的速率都没有差异(46/310 [14.8%]与35/298 [11.7%]) ,分别为P> 0.05)。此外,在20和5%的氧气张力下,胚胎的发育比例相似(18/298与27/310)。卵母细胞的核成熟(中期II),在精子头的浓缩,原核的形成,卵裂和胚胎发育方面,在大气(20%O-2; 12.4%[37/298])之间相似(P = 0.299)。并在体外受精(IVF)后在体外培养(IVC)的所有步骤中降低了氧气张力(5%O-2; 15.8%[49/310])。据我们所知,这是第一项证明犬胚可以使用低氧体外培养系统生产的研究。两种氧气张力(5%和20%)都导致相似的胚胎发育,因此对于犬卵母细胞的IVC是可行的。 (C)2013 Elsevier Inc.保留所有权利。

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