本文選題：牛胚胎 + 體外受精��； 參考：《吉林大學(xué)》2017年碩士論文

【摘要】：隨著牛畜產(chǎn)業(yè)的現(xiàn)代化發(fā)展,牛胚胎體外生產(chǎn)、胚胎冷凍、胚胎移植,已經(jīng)成為牛生產(chǎn)繁殖的重要技術(shù)手段。然而在胚胎體外生產(chǎn)與胚胎冷凍過程中,一些問題仍然不明確或存在爭(zhēng)議,從而導(dǎo)致體外胚胎生產(chǎn)效率低、胚胎冷凍成功率低等現(xiàn)象。活體采卵技術(shù)(Ovum Pick-up,OPU)的出現(xiàn),為牛胚胎體外生產(chǎn)提供了源源不斷的卵母細(xì)胞,更加促進(jìn)了這一產(chǎn)業(yè)的發(fā)展。但是,由于這一技術(shù)的引入以及現(xiàn)場(chǎng)實(shí)際條件的制約,使得傳統(tǒng)的實(shí)驗(yàn)室胚胎生產(chǎn)體系不再適用,需要進(jìn)行優(yōu)化調(diào)整。ε-聚賴氨酸(ε-Poly-L-lysine,ε-PL)具有抗菌性良好、有效濃度低、安全無毒、可生物降解、水溶性好、熱穩(wěn)定性好等優(yōu)點(diǎn),且一些研究表明,ε-PL對(duì)細(xì)胞膜以及核酸具有一定的保護(hù)作用,因此預(yù)計(jì)它可在胚胎冷凍中起到保護(hù)作用。1.本實(shí)驗(yàn)對(duì)牛胚胎體外生產(chǎn)體系進(jìn)行了適當(dāng)?shù)膬?yōu)化與調(diào)整,使其更加適用于現(xiàn)場(chǎng)胚胎生產(chǎn),研究表明:(1)與5%CO2、5%O2、90%N2的培養(yǎng)體系相比,5%CO2、95%空氣的培養(yǎng)體系可以取得更高的囊胚率(29.28%,7.29%);(2)當(dāng)培養(yǎng)液用量為每個(gè)胚胎5 uL時(shí),其囊胚率要高于每個(gè)胚胎10 uL與3 uL(43.52%,28.57%,40.11%);(3)當(dāng)胚胎培養(yǎng)數(shù)量為10個(gè)時(shí),其囊胚率要高于培養(yǎng)數(shù)量為5個(gè)與3個(gè),采用共培養(yǎng)體系可以提高囊胚率(43.52%,22.83%,16.09%,32.04%);(4)在IVM前保留全部卵丘細(xì)胞,IVF前去除適當(dāng)?shù)穆亚鸺?xì)胞,可以取得更高的囊胚率。2.實(shí)驗(yàn)探討了ε-PL在牛胚胎玻璃化冷凍中的保護(hù)作用,研究表明:(1)ε-PL不可單獨(dú)作為冷凍保護(hù)劑,進(jìn)行胚胎冷凍;(2)傳統(tǒng)玻璃化冷凍保護(hù)劑在添加ε-PL后,可以取得更加的保護(hù)效果,在0 g/L、10 g/L、20 g/L、30 g/L四個(gè)濃度中,20 g/L的冷凍效果最佳。(3)當(dāng)冷凍液緩沖時(shí)間為7.5 min時(shí),取得的胚胎冷凍效果要優(yōu)于5 min、10 min。實(shí)驗(yàn)結(jié)果表明,ε-PL對(duì)牛胚胎玻璃化冷凍具有保護(hù)作用.
[Abstract]:With the modernization of cattle and livestock industry, bovine embryo production in vitro, embryo freezing and embryo transfer has become an important technical means for cattle production and reproduction. However, in the process of in vitro embryo production and embryo freezing, some problems are still unclear or controversial, which leads to the low efficiency of in vitro embryo production and the low success rate of embryo freezing. The emergence of Ovum Pick-upopu (Ovum Pick-upOPU) provides a continuous stream of oocytes for in vitro production of bovine embryos and promotes the development of the industry. However, due to the introduction of this technique and the restriction of the field conditions, the traditional laboratory embryo production system is no longer applicable and needs to be optimized and adjusted. 蔚 -Poly-L-lysine (蔚 -PL) has good antibacterial properties, low effective concentration, safe and non-toxic. Some studies have shown that 蔚 -PL has some protective effects on cell membrane and nucleic acid, so it is expected that 蔚 -PL can play a protective role in embryo freezing. The system of bovine embryo production in vitro was optimized and adjusted to make it more suitable for field embryo production. The results showed that the blastocyst rate was higher than that of 5CO _ 2N _ 2 and 5CO _ 2O _ 290N _ 2. A higher blastocyst rate could be obtained when the amount of culture medium was 5uL, and the blastocyst rate was higher than that of each embryo (10uL and 3uL43.52g 28.577.57) when the number of embryo culture was 10, the blastocyst rate was higher than that of 5uL) when the number of embryos cultured was 10, the blastocyst rate was higher. The blastocyst rate of the blastocyst was higher than the number of 5 and 3. The blastocyst rate could be increased by using co-culture system. The blastocyst rate was increased by 22.83% and 16.09% and 32.04% respectively) the proper cumulus cells could be removed before IVM was retained, and a higher blastocyst rate of 0.2 could be obtained. The protective effect of 蔚 -PL on vitrification of bovine embryos was studied. The results showed that 蔚 -PL could not be used as cryopreservation agent alone. At 0 g / L 10 g / L 20 g / L 30 g / L, 20 g / L, the best freezing effect is 20 g / L) when the buffer time of the solution is 7.5 min, the embryo freezing effect is better than that of 5 min or 10 min. The results showed that 蔚-PL had protective effect on vitrification of bovine embryos.
【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】：2017
【分類號(hào)】：S823

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 林峰;黃承俊;宋歡;孫守強(qiáng);王幸栓;李峰;梁景山;胡艷芳;侯發(fā)全;劉海軍;;受精液種類、卵丘細(xì)胞及卵巢保存溫度對(duì)奶牛體外受精效果的影響[J];天津農(nóng)業(yè)科學(xué);2016年07期

2 陳江凌;汪聰勇;王現(xiàn)軍;田全召;;河南省荷斯坦牛冷凍胚胎移植效果分析[J];中國(guó)奶牛;2016年02期

3 余柳枝;陸超;劉道軍;;基于聚賴氨酸和聚乙二醇星形嵌段共聚物的基因載體研究[J];汕頭大學(xué)醫(yī)學(xué)院學(xué)報(bào);2014年03期

4 張丹丹;顏云松;劉道軍;;基于星形聚賴氨酸的基因載體研究[J];汕頭大學(xué)醫(yī)學(xué)院學(xué)報(bào);2014年02期

5 韓偉;馮春濤;畢江華;陳龍;劉瀟;裴燕;余文莉;;添加阿拉伯半乳聚糖或聚蔗糖對(duì)牛取樣胚胎冷凍效果的影響[J];中國(guó)畜牧獸醫(yī);2013年08期

6 劉迎春;邢瓊;周平;紀(jì)冬梅;陳蓓麗;蘇天紅;章志國(guó);曹云霞;;低氧對(duì)體外受精-胚胎移植胚胎發(fā)育潛能的影響[J];生殖與避孕;2012年04期

7 王軍;孫博;呂文發(fā);;哺乳動(dòng)物胚胎冷凍保存研究進(jìn)展[J];安徽農(nóng)業(yè)科學(xué);2007年34期

8 張東榮,王正剛,毛忠貴;聚賴氨酸的研究進(jìn)展[J];氨基酸和生物資源;2005年02期

9 郭憲,陳學(xué)進(jìn);培養(yǎng)液體積對(duì)牛卵母細(xì)胞體外成熟的影響[J];中國(guó)畜牧雜志;2004年11期

10 于永生,常萬存,李青旺;培養(yǎng)液體積對(duì)牛卵泡卵母細(xì)胞IVM效果的影響[J];中國(guó)草食動(dòng)物;2003年06期

相關(guān)碩士學(xué)位論文 前1條

1 馬云;牛卵泡卵母細(xì)胞體外成熟、體外受精及受精卵體外培養(yǎng)的研究[D];西北農(nóng)林科技大學(xué);2001年

，

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