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在通过灿烂甲酚蓝试验筛选的青春期前山羊卵母细胞的成熟和胚胎培养过程中添加半胱胺对胚胎发育的影响。

Supplementation with cysteamine during maturation and embryo culture on embryo development of prepubertal goat oocytes selected by the brilliant cresyl blue test.

作者信息

Urdaneta Aixa, Jiménez-Macedo Ana-Raquel, Izquierdo Dolors, Paramio Maria-Teresa

机构信息

Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain.

出版信息

Zygote. 2003 Nov;11(4):347-54. doi: 10.1017/s0967199403002405.

Abstract

Our previous studies have shown that the addition of 100 mircroM cysteamine to the in vitro maturation (IVM) medium increased the embryo development of prepubertal goat oocytes. The aim of the present study was to evaluate the effect of adding different concentrations of cysteamine to the IVM medium and to the in vitro embryo culture medium (IVC) on the embryo development of prepubertal goat oocytes selected by the brilliant cresyl blue (BCB) test. Oocytes were exposed to BCB and classified as: oocytes with a blue cytoplasm or grown oocytes (BCB+) or oocytes without blue cytoplasm or growing oocytes (BCB-). In Experiment 1, oocytes were matured in a conventional IVM medium supplemented with 100 microM, 200 microM or 400 microM cysteamine. In Experiment 2, oocytes were matured with 400 microM cysteamine and following in vitro fertilization (IVF) were cultured in SOF medium supplemented with 50 microM and 100 microM cysteamine. In Experiment 1, BCB+ oocytes matured with 100 microM and 200 microM cysteamine showed higher normal fertilization and embryo development rates than BCB- oocytes. Oocytes matured with 400 microM cysteamine did not present these differences between BCB+ and BCB- oocytes. In Experiment 2, the addition of 50 microM and 100 microM cysteamine to culture medium did not affect the proportion of total embryos obtained from BCB+ oocytes (35.89% and 38.29%, respectively) but was significantly different in BCB- oocytes (34.23% and 29.04%, respectively, P < 0.05). In conclusion, the addition of 400 microM cysteamine to the IVM improved normal fertilization and embryo development of BCB- oocytes at the same rates as those obtained from BCB+ oocytes. The proportions of morulae plus blastocyst development were not affected by the treatments.

摘要

我们之前的研究表明,在体外成熟(IVM)培养基中添加100微摩尔半胱胺可提高青春期前山羊卵母细胞的胚胎发育率。本研究的目的是评估在IVM培养基和体外胚胎培养基(IVC)中添加不同浓度半胱胺对经灿烂甲酚蓝(BCB)试验筛选的青春期前山羊卵母细胞胚胎发育的影响。将卵母细胞用BCB处理并分类为:细胞质呈蓝色的卵母细胞或生长中的卵母细胞(BCB+),或细胞质无蓝色的卵母细胞或未生长的卵母细胞(BCB-)。在实验1中,卵母细胞在添加了100微摩尔、200微摩尔或400微摩尔半胱胺的传统IVM培养基中成熟。在实验2中,卵母细胞用400微摩尔半胱胺成熟,体外受精(IVF)后在添加了50微摩尔和100微摩尔半胱胺的SOF培养基中培养。在实验1中,用100微摩尔和200微摩尔半胱胺成熟的BCB+卵母细胞比BCB-卵母细胞显示出更高的正常受精率和胚胎发育率。用400微摩尔半胱胺成熟的卵母细胞在BCB+和BCB-卵母细胞之间未呈现这些差异。在实验2中,向培养基中添加50微摩尔和100微摩尔半胱胺对从BCB+卵母细胞获得的总胚胎比例(分别为35.89%和38.29%)没有影响,但在BCB-卵母细胞中差异显著(分别为34.23%和29.04%,P<0.05)。总之,在IVM中添加400微摩尔半胱胺可提高BCB-卵母细胞的正常受精率和胚胎发育率,其速率与从BCB+卵母细胞获得的相同。桑椹胚加囊胚发育的比例不受处理的影响。

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