本文選題：小鼠 + 超數(shù)排卵��； 參考：《沈陽農(nóng)業(yè)大學(xué)》2016年碩士論文

【摘要】：小鼠在人和動(dòng)物生物、生理、遺傳研究等方面應(yīng)用廣泛,在包括腫瘤、遺傳病等生物醫(yī)學(xué)領(lǐng)域中被視為理想的動(dòng)物模型。隨著人們對(duì)轉(zhuǎn)基因小鼠研究興趣日益濃厚,量產(chǎn)轉(zhuǎn)基因小鼠和對(duì)小鼠進(jìn)行保種及研究需求日益增多。超數(shù)排卵所使用的供體雌鼠周齡可對(duì)收集的卵母細(xì)胞或受精卵的質(zhì)量和數(shù)量造成影響。近年來,學(xué)者們?cè)谥荦g對(duì)小鼠繁殖力的影響方面,存在周齡分組較少,對(duì)12-28周齡小鼠的超數(shù)排卵效果對(duì)比試驗(yàn)少見報(bào)道。故本試驗(yàn)對(duì)小鼠周齡進(jìn)行詳細(xì)分組,旨在探討小鼠周齡對(duì)超數(shù)排卵效果及體外培養(yǎng)發(fā)育率的影響；在胚胎移植方面,小鼠胚胎移植后產(chǎn)仔率仍不理想。有關(guān)胚胎移植最佳胚胎數(shù)量及單、雙側(cè)的輸卵管和子宮移植方式對(duì)比試驗(yàn)鮮有報(bào)道,故本試驗(yàn)對(duì)移植胚胎數(shù)量、發(fā)育階段及部位進(jìn)行對(duì)比研究。本試驗(yàn)選擇昆明和C57BL/6小鼠為實(shí)驗(yàn)動(dòng)物。因?yàn)槔ッ餍∈筮m應(yīng)力強(qiáng),繁殖率及成活率高,而C57BL/6小鼠為國際通用實(shí)驗(yàn)動(dòng)物。本論文分二部分試驗(yàn),試驗(yàn)一以各周齡(4-28周齡,共13組的昆明雌鼠；4-24周齡,共11組的C57BL/6雌鼠)小鼠為實(shí)驗(yàn)動(dòng)物。對(duì)所有試驗(yàn)小鼠進(jìn)行超數(shù)排卵處理,合籠檢栓,然后收集1-細(xì)胞和2-細(xì)胞胚胎進(jìn)行計(jì)數(shù),挑選質(zhì)量良好的胚胎進(jìn)行體外培養(yǎng),觀察受精卵的體外發(fā)育情況。試驗(yàn)二以昆明和C57BL/6雌鼠為實(shí)驗(yàn)動(dòng)物,研究胚胎在移植數(shù)量(10,15和20)、發(fā)育階段(1-細(xì)胞、2-細(xì)胞及囊胚)及部位(單雙側(cè)輸卵管及子宮)方面對(duì)胚胎移植效果的影響。試驗(yàn)一結(jié)果表明,收集的8-10周齡昆明(52.50-58.75枚)和C57BL/6(17.50-24.25枚)雌鼠可用受精卵數(shù)顯著高于其他周齡組；2-細(xì)胞胚胎體外培養(yǎng)發(fā)育率顯著高于1-細(xì)胞期胚胎；將收集到的同周齡不同品系的封閉群昆明和近交系C57BL/6小鼠進(jìn)行比對(duì)之后,兩者表現(xiàn)為極顯著差異。試驗(yàn)二結(jié)果表明,假孕昆明雌鼠移植20枚1-細(xì)胞胚胎的產(chǎn)仔率差異極顯著于10和15枚,其中20枚的產(chǎn)仔率顯著高于10和15枚(昆明組63.75%vs 40.00%,33.33%；C57BL/6組62.50%vs 45.00%,46.67%)；昆明和C57BL/6小鼠的1-細(xì)胞期胚胎組移植后的產(chǎn)仔率與移植2-細(xì)胞期胚胎組均差異顯著,其中1-細(xì)胞期胚胎移植組顯著高于2-細(xì)胞期胚胎組(昆明組67.50% vs 48.75%;C57BL/6組65.00% vs 48.75%),與移植囊胚組差異均不顯著,2-細(xì)胞期胚胎與囊胚組差異不顯著；昆明與C57BL/6小鼠胚胎移植于假孕昆明雌鼠的不同部位(單側(cè)輸卵管、雙側(cè)輸卵管、單側(cè)子宮或雙側(cè)子宮)的產(chǎn)仔率之間均差異不顯著；在移植數(shù)量、胚胎發(fā)育階段及移植部位相同的情況下,不同品系的昆明和C57BL/6小鼠的產(chǎn)仔率差異均不顯著。本試驗(yàn)以昆明和C57BL/6小鼠為實(shí)驗(yàn)動(dòng)物,探討了周齡對(duì)超數(shù)排卵及胚胎體外發(fā)育的影響,找到了昆明和C57BL/6小鼠最佳超數(shù)排卵的周齡；通過對(duì)胚胎移植產(chǎn)仔率影響因素的研究(移植胚胎數(shù)量、發(fā)育階段及移植部位),找到可顯著提高胚胎移植產(chǎn)仔率的方法,為今后有關(guān)小鼠胚胎移植技術(shù)提供參考。
[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.
【學(xué)位授予單位】：沈陽農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S814

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