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在牛激活素A存在的情况下,牛卵母细胞的体外成熟不会影响胚胎数量。

In vitro maturation of bovine oocytes in the presence of bovine activin A does not affect the number of embryos.

作者信息

Izadyar F, Zeinstra E, Colenbrander B, Vanderstichele H M, Bevers M M

机构信息

Department of Herd Health and Reproduction, Faculty of Veterinary Medicine, University of Utrecht, Netherlands.

出版信息

Anim Reprod Sci. 1996 Dec 2;45(1-2):37-45. doi: 10.1016/s0378-4320(96)01574-6.

Abstract

This study was carried out to investigate whether bovine recombinant activin A present during the in vitro maturation (IVM) of cumulus enclosed bovine oocytes affects the proportion of embryos that develop to the blastocyst stage. In addition, the effect of the presence of activin A during maturation and during embryo culture was studied. Therefore, bovine cumulus oocyte complexes were matured at 39 degrees C in a humidified atmosphere with 5% CO2 in air for 24 h in: (1) culture medium M199 supplemented with 10% foetal calf serum (FCS), luteinising hormone (LH) and follicle-stimulating hormone (FSH) and 10 ng ml-1 activin A; (2) M199 without FCS but supplemented with LH and FSH and 10 ng ml-1 activin A; (3) M199 without FCS, LH and FSH but supplemented with 10 ng ml-1 activin A. Cultures without activin served as controls. After IVF the embryos were cultured in M199 supplemented with 10% FCS on a monolayer of buffalo rat liver (BRL) cells. For the second part of the study, COCs were matured in vitro in M199 supplemented with LH and FSH and 10 ng ml-1 activin A, fertilized in vitro and the embryos were cultured (1) on a monolayer of BRL cells in M199 supplemented with 10% FCS and 10 ng ml-1 of activin A, and (2) in droplets of serum free BRL-conditioned medium supplemented with 10 ng ml-1 activin A. IVM in the presence of LH, FSH and 10 ng ml-1 activin A did not change the proportion of blastocysts present at Day 9 or the proportion of hatched blastocyst at Day 11. Activin present during maturation in the absence of serum and gonadotrophic hormones also did not alter the proportion of blastocysts or hatched blastocysts. In vitro culture of embryos on BRL cells or in BRL-conditioned medium in the presence of activin had no effect on embryonic development. It is concluded that IVM in the presence of bovine activin A has no effect on subsequent embryonic development.

摘要

本研究旨在调查卵丘包裹的牛卵母细胞体外成熟(IVM)过程中存在的牛重组激活素A是否会影响发育至囊胚阶段的胚胎比例。此外,还研究了激活素A在成熟过程和胚胎培养过程中存在时的影响。因此,将牛卵丘卵母细胞复合体在39℃、含5%二氧化碳的湿润空气中于以下培养基中成熟24小时:(1)添加10%胎牛血清(FCS)、促黄体生成素(LH)和促卵泡激素(FSH)以及10 ng/ml激活素A的M199培养基;(2)不含FCS但添加LH、FSH和10 ng/ml激活素A的M199培养基;(3)不含FCS、LH和FSH但添加10 ng/ml激活素A的M199培养基。不添加激活素的培养物作为对照。体外受精后,将胚胎在添加10% FCS的M199培养基中于水牛大鼠肝(BRL)细胞单层上培养。对于研究的第二部分,将卵丘卵母细胞复合体在添加LH、FSH和10 ng/ml激活素A的M199培养基中体外成熟,体外受精,然后将胚胎培养:(1)在添加10% FCS和10 ng/ml激活素A的M199培养基中的BRL细胞单层上培养;(2)在添加10 ng/ml激活素A的无血清BRL条件培养基液滴中培养。在LH、FSH和10 ng/ml激活素A存在下进行IVM,并不会改变第9天囊胚的比例或第11天孵化囊胚的比例。在无血清和促性腺激素的情况下成熟过程中存在的激活素也不会改变囊胚或孵化囊胚的比例。在激活素存在的情况下,在BRL细胞上或BRL条件培养基中对胚胎进行体外培养对胚胎发育没有影响。得出的结论是,在牛激活素A存在的情况下进行IVM对随后的胚胎发育没有影响。

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