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用于改善牛体外胚胎生产的模拟生理性卵母细胞成熟系统的评估。

Evaluation of the simulated physiological oocyte maturation system for improving bovine in vitro embryo production.

作者信息

Guimarães A L S, Pereira S A, Leme L O, Dode M A N

机构信息

School of Agriculture and Veterinary Medicine, University of Brasília, Brasília, Distrito Federal Brazil.

出版信息

Theriogenology. 2015 Jan 1;83(1):52-7. doi: 10.1016/j.theriogenology.2014.07.042.

Abstract

The aim of this study was to test the simulated physiological oocyte maturation (SPOM)- adapted system during bovine oocyte maturation to improve embryo development. Oocytes were obtained from follicles of 3 to 8 mm in diameter that were aspirated from ovaries obtained from a slaughterhouse. To verify the effect of the maturation system on in vitro embryo production, the cleavage, blastocyst rates on Days 7 and 8, embryo size, and total cell number were evaluated. The resulting data on embryo development were analyzed by the chi-square test, whereas data on embryo size and total cell number were analyzed by the Kruskal-Wallis test. First, the SPOM system principle was tested in our IVM system, in which 0.01 IU/mL of purified FSH and 10% of fetal calf serum were used during maturation. However, the cleavage and blastocyst rates on Days 7 and 8 were drastically reduced compared with those of the control group (P < 0.05). Increasing the dose of purified FSH to 0.1 IU/mL in the SPOM-adapted system did not affect (P > 0.05) embryo production, which remained lower than that of the control group. When less competent oocytes obtained from 1 to 3 mm follicles were used, the SPOM-adapted system was also unable to improve embryo production. To make the adapted system as similar as possible to the reported system, recombinant FSH was associated with BSA during maturation and embryo culture was performed under low oxygen tension conditions. Nevertheless, a reduction (P < 0.05) in the blastocyst rates was also observed, whereas the size and total cell number were similar to those of the control group (P > 0.05). It can be concluded that an SPOM-adapted system used under different culture conditions does not improve in vitro embryo development.

摘要

本研究的目的是在牛卵母细胞成熟过程中测试模拟生理卵母细胞成熟(SPOM)适配系统,以改善胚胎发育。卵母细胞从屠宰场获得的卵巢中抽吸的直径为3至8毫米的卵泡中获取。为了验证成熟系统对体外胚胎生产的影响,评估了第7天和第8天的卵裂率、囊胚率、胚胎大小和总细胞数。胚胎发育的结果数据采用卡方检验进行分析,而胚胎大小和总细胞数的数据采用Kruskal-Wallis检验进行分析。首先,在我们的体外成熟(IVM)系统中测试了SPOM系统原理,在成熟过程中使用了0.01 IU/mL的纯化促卵泡素(FSH)和10%的胎牛血清。然而,与对照组相比,第7天和第8天的卵裂率和囊胚率大幅降低(P < 0.05)。在SPOM适配系统中将纯化FSH的剂量增加到0.1 IU/mL对胚胎生产没有影响(P > 0.05),其仍低于对照组。当使用从1至3毫米卵泡中获得的能力较差的卵母细胞时,SPOM适配系统也无法提高胚胎生产。为了使适配系统尽可能与报道的系统相似,在成熟过程中将重组FSH与牛血清白蛋白(BSA)结合,并在低氧张力条件下进行胚胎培养。然而,也观察到囊胚率降低(P < 0.05),而大小和总细胞数与对照组相似(P > 0.05)。可以得出结论,在不同培养条件下使用的SPOM适配系统并不能改善体外胚胎发育。

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