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在不同成熟时间的体外成熟过程中添加咖啡因对犬卵母细胞发育能力的改善

Improvement of the developmental competence of canine oocyte using caffeine supplementation during IVM at different maturation time.

作者信息

Fathi Mohamed, Salama A, Badr Magdy R

机构信息

Department of Theriogenology,Faculty of Veterinary Medicine,Cairo University,Giza,Egypt.

Department of AI and ET,Animal Reproduction Research Institute,Agriculture Research Center,Giza,Egypt.

出版信息

Zygote. 2018 Apr;26(2):162-167. doi: 10.1017/S0967199418000059. Epub 2018 May 25.

Abstract

SummaryThe aim of the current study was to investigate the effect of caffeine supplementation during in vitro maturation (IVM) for different maturation times on the developmental potential of canine oocytes recovered from ovariohysterectomized bitches. The recovered cumulus-oocytes complexes were in vitro matured for 72 h. Here, 10 mM caffeine was added to the maturation medium for different incubation times (caffeine from 0-72 h maturation, caffeine for the first 24 h of maturation only, caffeine addition from 24 to 48 h maturation time, caffeine addition from 48 to 72 h maturation or in caffeine-free medium, control group). The matured oocytes were in vitro fertilized using frozen-thawed spermatozoa. The presumptive zygotes were in vitro cultured in synthetic oviductal fluid medium for 5 days. The results showed that both maturation and fertilization rates were significantly higher (P ˂ 0.05) using caffeine-treated medium for the first 24 h of maturation compared with the control and other two groups of caffeine treatment (from 24 to 48 h and from 48 to 72 h), whereas use of caffeine-treated medium for a 0-72 h incubation time did not affect these rates (P > 0.05). Interestingly, the matured oocytes in caffeine-supplemented medium for the first 24 h or from 0-72 h showed a significant (P ˂ 0.05) increase in the total number of cleaved embryos compared with the control group. In conclusion, supplementation of the maturation medium with 10 mM caffeine for the first 24 h of maturation or during the whole maturation time (0-72 h) improved nuclear maturation and subsequent embryo development preimplantation following in vitro fertilization.

摘要

摘要

本研究的目的是调查在体外成熟(IVM)不同成熟时间补充咖啡因对从卵巢子宫切除母犬回收的犬卵母细胞发育潜力的影响。回收的卵丘-卵母细胞复合体在体外成熟72小时。在此,将10 mM咖啡因添加到成熟培养基中,用于不同的孵育时间(从0至72小时成熟阶段添加咖啡因、仅在成熟的前24小时添加咖啡因、在24至48小时成熟阶段添加咖啡因、在48至72小时成熟阶段添加咖啡因或在无咖啡因培养基中,即对照组)。成熟的卵母细胞使用冻融精子进行体外受精。假定的受精卵在合成输卵管液培养基中体外培养5天。结果显示,与对照组和其他两组咖啡因处理组(24至48小时和48至72小时)相比,在成熟的前24小时使用咖啡因处理的培养基时,成熟率和受精率均显著更高(P<0.05),而在0至72小时孵育时间使用咖啡因处理的培养基并不影响这些比率(P>0.05)。有趣的是,与对照组相比,在成熟的前24小时或0至72小时补充咖啡因的培养基中的成熟卵母细胞,其分裂胚胎总数显著增加(P<0.05)。总之,在成熟的前24小时或整个成熟时间(0至72小时)向成熟培养基中补充10 mM咖啡因可改善核成熟以及体外受精后植入前的后续胚胎发育。

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