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The effect of temperature during liquid storage of in vitro-matured bovine oocytes on subsequent embryo development

机译:体外成熟牛卵母细胞液体储存过程中温度对随后胚胎发育的影响

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The aim of the present study was to optimize the temperature for the temporal storage of matured bovine oocytes. In vitro-matured bovine oocytes were preserved in HEPES-buffered TCM199 medium supplemented with 10% newborn calf serum at different temperatures (4 degrees C, 15 degrees C, 25 degrees C, and 38.5 degrees C) for 20 hours. Embryo development and blastocyst quality after in vitro fertilization, cytoplasmic ATP and glutathione levels in oocytes, and the frequency of apoptotic oocytes were compared among storage groups and a control group without storage. Among the storage groups, those at 25 degrees C and 38.5 degrees C showed the highest rates of blastocyst development (19.3% and 24.5%, respectively) compared with those stored at 4 degrees C and 15 degrees C (8.5% and 14.9%, respectively); however, blastocyst formation rates in all storage groups were lower than that in the control group (39.8%; P 0.05). Storage at 38.5 degrees C and 15 degrees C was associated with reduced cell numbers in resultant blastocysts compared with the control and the 25 degrees C storage groups. Storage at 4 degrees C reduced metabolic activity of oocytes characterized by their lower ATP levels compared with the other groups. Storage for 20 hours significantly reduced the glutathione content in oocytes in all groups in a similar manner, irrespective of the temperature. Storage at 4 degrees C or 15 degrees C but not at 25 degrees C and 38.5 degrees C significantly increased the percentage of apoptotic oocytes compared with the control group. In conclusion, 25 degrees C was found to be the most suitable temperature for the temporal storage of matured bovine oocytes regarding both the developmental competence of oocytes and the quality of resultant blastocysts. (c) 2016 Elsevier Inc. All rights reserved.
机译:本研究的目的是优化成熟牛卵母细胞暂时储存的温度。将体外成熟的牛卵母细胞在补充有10%新生小牛血清的HEPES缓冲TCM199培养基中在不同温度(4摄氏度,15摄氏度,25摄氏度和38.5摄氏度)下保存20小时。比较贮藏组和无贮藏组的体外受精后的胚胎发育和胚泡质量,卵母细胞中细胞质ATP和谷胱甘肽水平以及凋亡卵母细胞的频率。在储存组中,与在4摄氏度和15摄氏度(分别为8.5%和14.9%)储存的相比,在25摄氏度和38.5摄氏度的储存组中胚泡发育率最高(分别为19.3%和24.5%)。 );然而,所有贮藏组的囊胚形成率均低于对照组(39.8%; P <0.05)。与对照组和25摄氏度的储存组相比,在38.5摄氏度和15摄氏度的储存条件下,胚泡的细胞数量减少。与其他组相比,在4°C下储存会降低卵母细胞的代谢活性,卵母细胞的ATP水平较低。储存20小时,无论温度如何,所有组的卵母细胞中谷胱甘肽的含量均以类似的方式降低。与对照组相比,在4摄氏度或15摄氏度而不是25摄氏度和38.5摄氏度的条件下进行储存显着增加了凋亡卵母细胞的百分比。总之,就卵母细胞的发育能力和所产生的胚泡的质量而言,发现25℃是最适合暂时储存成熟牛卵母细胞的温度。 (c)2016 Elsevier Inc.保留所有权利。

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