【摘要】：卵母細(xì)胞體外成熟（in vitro maturation,IVM）和體細(xì)胞克隆（somatic cell nuclear transfer,SCNT）技術(shù)在動(dòng)物繁殖領(lǐng)域的應(yīng)用已經(jīng)十分廣泛，但在實(shí)際生產(chǎn)和試驗(yàn)研究中仍存在許多問(wèn)題。如IVM卵母細(xì)胞質(zhì)量不如體內(nèi)自然正常發(fā)育的卵母細(xì)胞，且其胚胎發(fā)育能力也不如體內(nèi)胚胎；SCNT胚胎的發(fā)育及克隆動(dòng)物生產(chǎn)效率低下，克隆胚胎妊娠率低、克隆胚胎流產(chǎn)率高等因素導(dǎo)致克隆犢牛出生率低，出生的克隆牛存活率也低，且大多數(shù)都有異常等。本試驗(yàn)研究了維生素C（Vtiamin C,Vc）對(duì)卵母細(xì)胞成熟及胚胎發(fā)育的影響及影響體細(xì)胞克隆牛生產(chǎn)效率的因素。結(jié)果表明：（1）卵母細(xì)胞成熟液中添加濃度為50μg/mL的Vc，可以提高卵母細(xì)胞的體外成熟率；成熟液中添加不同濃度的Vc體外成熟培養(yǎng)卵母細(xì)胞，各組間孤雌胚胎的卵裂率和囊胚率沒(méi)有顯著差異；胚胎培養(yǎng)液中添加不同濃度的Vc培養(yǎng)體外受精胚胎，20μg/mL Vc組能顯著提高胚胎的卵裂率，各組間的囊胚率無(wú)顯著差異，多能性相關(guān)基因的表達(dá)有所差異；成熟液和培養(yǎng)液中同時(shí)添加Vc能夠提高體外受精胚胎的卵裂率，不影響囊胚形成率和囊胚的總細(xì)胞數(shù)，但促進(jìn)了多能性相關(guān)基因的表達(dá)；（2）體細(xì)胞克隆牛生產(chǎn)效率的影響因素方面，移植胚胎的數(shù)量以2枚胚胎最適宜；胚胎移植的季節(jié)應(yīng)選在春、秋兩季；受體牛應(yīng)選擇黃體等級(jí)為A、B級(jí)的母牛作為受體。本試驗(yàn)從卵母細(xì)胞的成熟培養(yǎng)、胚胎的體外發(fā)育、體細(xì)胞克隆牛生產(chǎn)等方面，系統(tǒng)研究了Vc對(duì)卵母細(xì)胞成熟和胚胎發(fā)育的作用，以及影響克隆牛生產(chǎn)效率的多種因素，對(duì)卵母細(xì)胞成熟和體細(xì)胞克隆進(jìn)行了有益的探討，為卵母細(xì)胞IVM和克隆牛生產(chǎn)研究提供了有利的依據(jù)。
[Abstract]:Oocyte maturation in vitro (in vitro maturation (in vitro) and somatic cell cloning (somatic cell nuclear transfer (somatic cell nuclear) have been widely used in animal reproduction, but there are still many problems in practical production and experimental research. For example, the quality of IVM oocytes is inferior to that of naturally developed oocytes in vivo, and its embryonic development ability is not as good as the development of IVM embryos in vivo and the production efficiency of cloned animals, and the pregnancy rate of cloned embryos is low. The high abortion rate of cloned embryos led to the low birth rate of cloned calves and the low survival rate of cloned cattle, and most of them were abnormal. The effects of vitamin C (Vtiamin C) on oocyte maturation and embryo development and the factors influencing the productivity of somatic cloned cattle were studied. The results showed that: (1) the rate of oocyte maturation could be increased by adding 50 渭 g/mL Vc to the maturation fluid of oocytes, and different concentrations of VC were added to the maturation fluid to culture oocytes. The cleavage rate and blastocyst rate of parthenogenetic embryos were not significantly different among different groups, but the blastocyst rate of 20 渭 g/mL VC group could significantly increase the blastocyst rate. The expression of pluripotent related genes was different, the cleavage rate of IVF embryos was increased by VC addition in maturation medium and culture medium, and the blastocyst formation rate and the total cell number of blastocyst were not affected, but the expression of pluripotent related genes was promoted. (2) in terms of the factors affecting the productivity of somatic cloned cattle, the number of embryos transferred was the most suitable, the transfer season should be in spring and autumn, and the recipient cattle should choose the cow with luteal grade as the receptor. In this study, the effects of VC on oocyte maturation and embryo development, embryo development in vitro and somatic cloned cattle production were studied systematically. The study of oocyte maturation and somatic cell cloning provided a useful basis for the study of oocyte IVM and cloned cattle production.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S823

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