维生素C对牛早期胚胎发育影响及体细胞克隆牛影响因素研究
发布时间：2018-07-26 20:16
【摘要】：卵母细胞体外成熟（in vitro maturation,IVM）和体细胞克隆（somatic cell nuclear transfer,SCNT）技术在动物繁殖领域的应用已经十分广泛，但在实际生产和试验研究中仍存在许多问题。如IVM卵母细胞质量不如体内自然正常发育的卵母细胞，且其胚胎发育能力也不如体内胚胎；SCNT胚胎的发育及克隆动物生产效率低下，克隆胚胎妊娠率低、克隆胚胎流产率高等因素导致克隆犊牛出生率低，出生的克隆牛存活率也低，且大多数都有异常等。本试验研究了维生素C（Vtiamin C,Vc）对卵母细胞成熟及胚胎发育的影响及影响体细胞克隆牛生产效率的因素。结果表明：（1）卵母细胞成熟液中添加浓度为50μg/mL的Vc，可以提高卵母细胞的体外成熟率；成熟液中添加不同浓度的Vc体外成熟培养卵母细胞，各组间孤雌胚胎的卵裂率和囊胚率没有显著差异；胚胎培养液中添加不同浓度的Vc培养体外受精胚胎，20μg/mL Vc组能显著提高胚胎的卵裂率，各组间的囊胚率无显著差异，多能性相关基因的表达有所差异；成熟液和培养液中同时添加Vc能够提高体外受精胚胎的卵裂率，不影响囊胚形成率和囊胚的总细胞数，但促进了多能性相关基因的表达；（2）体细胞克隆牛生产效率的影响因素方面，移植胚胎的数量以2枚胚胎最适宜；胚胎移植的季节应选在春、秋两季；受体牛应选择黄体等级为A、B级的母牛作为受体。本试验从卵母细胞的成熟培养、胚胎的体外发育、体细胞克隆牛生产等方面，系统研究了Vc对卵母细胞成熟和胚胎发育的作用，以及影响克隆牛生产效率的多种因素，对卵母细胞成熟和体细胞克隆进行了有益的探讨，为卵母细胞IVM和克隆牛生产研究提供了有利的依据。
[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.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S823
本文编号：2147170
[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.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S823
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