EGF和GSH对牛卵母细胞体外受精的影响

Effect of EGF and GSH on the in vitro Fertilization inBovine Oocyte

研究了牛卵泡卵母细胞体外受精时向成熟液中加入EGF(表皮生长因子),受精液中加入GSH(谷胱甘肽)对受精率和囊胚率的影响。成熟液以M-199为基础液并加入一定浓度的促性腺激素和OCS,作为成熟液的对照组,试验组加入不同浓度的EGF。受精液以改良TALP为基础液并加入一定浓度的BSA、肝素和咖啡因,作为受精液的对照组,试验组加入不同浓度的GSH。早期胚胎培养液为SOFaa。在成熟液中加入10ng/mlEGF和50ng/mlEGF组时,试验组与对照组的受精率分别为76.00%(114/150)、76.71%(112/146)和60.70%(85/140);囊胚率分别为23.68%(27/114)、24.11%(27/112)和17.64%(15/85);在受精液中加入1mMGSH和10mMGSH组时,试验组与对照组的受精率分别为71.66%(86/120)、63.28%(81/128)和76.00%(114/150);囊胚率分别为34.88%(30/86)、24.69%(20/81)和23.68%(27/114)。结果表明,成熟液中加入EGF对牛卵母细胞的体外成熟和受精后的早期胚胎的发育都有促进作用,在一定范围内增加EGF浓度,不会提高受精率和囊胚率;受精液中加入一定浓度GSH能够提高受精率和囊胚率,但是在高浓度的GSH时则影响受精率。

In present study the effects of EGF and GSH on the sheep in vitro fertilization of bovine were investigated, using M-199 as maturation control medium supplied with 5% OCS, using TALP medium as fertilization control medium supplied with 6%BSA, 5iu/ml heparine and 5mM caffine. The culture medium is SOFaa. When 10ng/mlEGF and 50ng/mlEGF is used in maturation medium, the the fertilization rate of EGF and control are 76.00%(114/150), 76.71% (112/146) and 60.70%(85/140),the blastcyst rate are 23.68%(27/114), 24.69%(20/81)and17.64%(15/85). The fertilization medium is TALP supplied with 6%BSA, 5iu/ml heparine and 5mM caffine. When 1mM GSH and 10mM is added, the fertilization rate of GSH and control is 71.66%(86/120), 63.28%(81/128) and 76.00%(114/150); the blastcyst rate is 34.88% (30/86), 24.69%(20/81)and 23.68%(27/114). The results indicate that M-199 supplement with 5% OCS and 10ng/mlEGF or 50ng/mlEGF can promote in vitro cleavage and development of bovine oocyte, but the higher concentration has no better effect than the lower concentration. At the same time, the fertilization medium added with 1mMGSH can promote in vitro development of bovine oocyte, without affect fertilization rate. But with the higher concentration of GSH, it cannot increase the rate of fertilization and blastcyst.