不同品系小鼠超数排卵与胚胎移植研究

发布时间：2018-05-31 16:43

本文选题：小鼠 + 超数排卵　；参考：《沈阳农业大学》2016年硕士论文

【摘要】：小鼠在人和动物生物、生理、遗传研究等方面应用广泛,在包括肿瘤、遗传病等生物医学领域中被视为理想的动物模型。随着人们对转基因小鼠研究兴趣日益浓厚,量产转基因小鼠和对小鼠进行保种及研究需求日益增多。超数排卵所使用的供体雌鼠周龄可对收集的卵母细胞或受精卵的质量和数量造成影响。近年来,学者们在周龄对小鼠繁殖力的影响方面,存在周龄分组较少,对12-28周龄小鼠的超数排卵效果对比试验少见报道。故本试验对小鼠周龄进行详细分组,旨在探讨小鼠周龄对超数排卵效果及体外培养发育率的影响；在胚胎移植方面,小鼠胚胎移植后产仔率仍不理想。有关胚胎移植最佳胚胎数量及单、双侧的输卵管和子宫移植方式对比试验鲜有报道,故本试验对移植胚胎数量、发育阶段及部位进行对比研究。本试验选择昆明和C57BL/6小鼠为实验动物。因为昆明小鼠适应力强,繁殖率及成活率高,而C57BL/6小鼠为国际通用实验动物。本论文分二部分试验,试验一以各周龄(4-28周龄,共13组的昆明雌鼠；4-24周龄,共11组的C57BL/6雌鼠)小鼠为实验动物。对所有试验小鼠进行超数排卵处理,合笼检栓,然后收集1-细胞和2-细胞胚胎进行计数,挑选质量良好的胚胎进行体外培养,观察受精卵的体外发育情况。试验二以昆明和C57BL/6雌鼠为实验动物,研究胚胎在移植数量(10,15和20)、发育阶段(1-细胞、2-细胞及囊胚)及部位(单双侧输卵管及子宫)方面对胚胎移植效果的影响。试验一结果表明,收集的8-10周龄昆明(52.50-58.75枚)和C57BL/6(17.50-24.25枚)雌鼠可用受精卵数显著高于其他周龄组；2-细胞胚胎体外培养发育率显著高于1-细胞期胚胎；将收集到的同周龄不同品系的封闭群昆明和近交系C57BL/6小鼠进行比对之后,两者表现为极显著差异。试验二结果表明,假孕昆明雌鼠移植20枚1-细胞胚胎的产仔率差异极显著于10和15枚,其中20枚的产仔率显著高于10和15枚(昆明组63.75%vs 40.00%,33.33%；C57BL/6组62.50%vs 45.00%,46.67%)；昆明和C57BL/6小鼠的1-细胞期胚胎组移植后的产仔率与移植2-细胞期胚胎组均差异显著,其中1-细胞期胚胎移植组显著高于2-细胞期胚胎组(昆明组67.50% vs 48.75%;C57BL/6组65.00% vs 48.75%),与移植囊胚组差异均不显著,2-细胞期胚胎与囊胚组差异不显著；昆明与C57BL/6小鼠胚胎移植于假孕昆明雌鼠的不同部位(单侧输卵管、双侧输卵管、单侧子宫或双侧子宫)的产仔率之间均差异不显著；在移植数量、胚胎发育阶段及移植部位相同的情况下,不同品系的昆明和C57BL/6小鼠的产仔率差异均不显著。本试验以昆明和C57BL/6小鼠为实验动物,探讨了周龄对超数排卵及胚胎体外发育的影响,找到了昆明和C57BL/6小鼠最佳超数排卵的周龄；通过对胚胎移植产仔率影响因素的研究(移植胚胎数量、发育阶段及移植部位),找到可显著提高胚胎移植产仔率的方法,为今后有关小鼠胚胎移植技术提供参考。
[Abstract]:Mice are widely used in human and animal biology, physiology, genetic research and other fields. It is regarded as an ideal animal model in biomedicine, including tumor and hereditary disease. With the increasing interest in transgenic mice, the demand for breeding and Research on transgenic mice and mice is increasing. Superovulation is used. The weeks of age in the donor mice can affect the quality and quantity of the oocyte or the fertilized egg. In recent years, there are few weeks of age grouping in the influence of the week age on the reproductive capacity of mice. This is a rare report on the comparison test of the superovulation effect of 12-28 weeks old mice. The effect of the age of mice on the effect of superovulation and the growth rate of the culture in vitro was discussed. In the embryo transplantation, the birth rate of the mouse embryo was still not ideal. The number of best embryos and the single, bilateral oviduct and uterine transplants were rarely reported. In this experiment, Kunming and C57BL/6 mice were selected as experimental animals. Because Kunming mice were resilient, reproductive rate and survival rate were high, and C57BL/6 mice were international common experimental animals. This paper was divided into two parts. The experiment was divided into two parts of each week (4-28 weeks of age, 13 groups of Kunming female rats; 4-24 weeks old, 11 groups of C57BL/6 female mice) Rats were treated as experimental animals. All the mice were treated with superovulation, combined with cage suppository, and then collected 1- and 2- cell embryos to count, select good embryos in vitro and observe the in vitro development of the fertilized eggs. Experiment two, Kunming and C57BL/6 female rats were used as experimental animals, and the number of embryos in transplantation (10,15 and 20) was studied. The effects of developmental stages (1- cells, 2- cells and blastocysts) and sites (single bilateral oviduct and uterus) on the effect of embryo transfer. Experimental results showed that the number of fertilized eggs available for 8-10 weeks old Kunming (52.50-58.75) and C57BL/6 (17.50-24.25) female mice was significantly higher than that of other weeks of age; the growth rate of 2- cell embryos in vitro was significantly higher than that of other weeks. Compared with the closed group Kunming and the inbred C57BL/6 mice of different lines of the same week age, the 1- cell stage embryos were significantly different. The results of test two showed that the birth rate of 20 1- cell embryos transplanted from female rats in pseudo pregnancy was significantly higher than that of 10 and 15, of which 20 had a significantly higher birth rate. 10 and 15 (Kunming group 63.75%vs 40%, 33.33%; C57BL/6 group 62.50%vs 45%, 46.67%); the birth rate of the 1- cell stage embryo group in Kunming and C57BL/6 mice was significantly different from that of the transplanted 2- cell stage embryo group, and the 1- cell stage embryo transplantation group was significantly higher than the 2- cell stage embryo group (67.50% vs 48.75% in Kunming group; C57BL/6 group 65%). Vs 48.75%), there was no significant difference between the transplanted blastocyst group and the 2- cell stage embryo and the blastocyst group, and there was no significant difference between the Kunming and C57BL/6 mouse embryos in the different parts of the female mice (unilateral fallopian tube, bilateral fallopian tube, unilateral uterus or bilateral uterus) in the female mice of pseudo pregnancy; the number of transplanted embryos and the embryonic development stage were not significant. The birth rates of Kunming and C57BL/6 mice in different strains were not significant. In this experiment, the effects of week age on Superovulation and in vitro development of embryos in Kunming and C57BL/6 mice were investigated, and the best weeks of superovulation in Kunming and C57BL/6 mice were found. The study of the factors affecting the rate of litter (the number of transplanted embryos, the stage of development and the site of transplantation), find a way to significantly increase the rate of embryo transfer, and provide a reference for the future technology of mouse embryo transfer.
【学位授予单位】：沈阳农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S814

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