本文選題：綿羊卵母細(xì)胞　切入點(diǎn)：體外成熟　出處：《河南農(nóng)業(yè)大學(xué)》2016年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：本實(shí)驗(yàn)重點(diǎn)研究了影響綿羊胞質(zhì)內(nèi)單精子注射(ICSI)中的因素,以期優(yōu)化綿羊ICSI的操作技術(shù)和體外成熟體系,為開(kāi)展發(fā)育機(jī)理研究及提高綿羊ICSI效率提供進(jìn)一步的技術(shù)支持。研究結(jié)果主要包括如下幾個(gè)方面:(1)不同季節(jié)對(duì)卵母細(xì)胞體外培養(yǎng)的影響。結(jié)果表明季節(jié)對(duì)卵母細(xì)胞的總數(shù)量沒(méi)有影響。春季和秋季獲得的可用卵母細(xì)胞數(shù)顯著高于夏季和冬季。就成熟率而言,春季、秋季和冬季差異不顯著。綜合回收率和成熟率,秋季是綿羊卵母細(xì)胞體外培養(yǎng)的最佳時(shí)節(jié),而夏季不適綿羊卵母細(xì)胞的采集培養(yǎng)。(2)不同成熟時(shí)間(20~22h、22~24h、24~26h、26~28h)對(duì)ICSI的影響。結(jié)果顯示卵母細(xì)胞的卵裂率隨成熟時(shí)間的增加而逐漸提高;卵母細(xì)胞成熟22~24 h桑囊率最高,之后開(kāi)始降低。因此,卵母細(xì)胞成熟22~24h是進(jìn)行ICSI的最佳時(shí)機(jī)。(3)高滲操作液對(duì)ICSI的影響,結(jié)果顯示高滲操作液能顯著提高操作成功率,但對(duì)卵裂率(43.9%、36.5%;P0.05)和桑囊率(41.4%、38.7%;P0.05)差異不顯著。(4)第一極體位置(6點(diǎn)和12點(diǎn))對(duì)ICSI的影響,結(jié)果發(fā)現(xiàn)極體位置對(duì)ICSI卵裂率(32.5%vs 34.5%,P0.05)和桑囊率(33.6%、33.3%;P0.05)都沒(méi)有顯著影響。(5)超聲波處理不同狀態(tài)精子(鮮精和凍精)對(duì)ICSI的影響。結(jié)果表明超聲波處理鮮精和凍精對(duì)卵母細(xì)胞的卵裂率(39.1%vs 37.4%,P0.05)和桑囊率(33.7%vs 35.1%,P0.05)均差異不顯著。(6)精子不同處理方式(NaOH和超聲波處理)對(duì)ICSI的影響,結(jié)果表明兩種處理方式對(duì)卵母細(xì)胞的卵裂率(37.4%vs 46.6%,P0.05)差異顯著;NaOH處理組的桑囊率(52.1%)差異顯著高于超聲波處理組(35.1%)。(7)精子共孵育對(duì)ICSI卵發(fā)育的影響。以NaOH處理精子ICSI作為對(duì)照,NaOH處理精子與質(zhì)粒共孵育后對(duì)ICSI胚的卵裂率(37.9%vs 46.6%,P0.05)和桑囊率(45.3%vs52.1%,P0.05)的影響差異不顯著。(8)非共培養(yǎng)和卵丘細(xì)胞共培養(yǎng)系統(tǒng)對(duì)ICSI胚的影響。結(jié)果顯示共培養(yǎng)和非共培養(yǎng)對(duì)卵裂率(41.7%vs 44.0%,P0.05)和桑囊率(41.8%vs 35.7%,P0.05)影響不顯著。
[Abstract]:In order to optimize the operation technology and in vitro maturation system of sheep ICSI, the factors affecting ICSI were studied. The results include the following aspects: 1) the effect of different seasons on the culture of oocytes in vitro. The results show that seasons affect oocytes fine. The number of available oocytes obtained in spring and autumn was significantly higher than that in summer and winter. There was no significant difference in spring, autumn and winter. The results showed that the cleavage rate of oocytes increased gradually with the increase of maturation time, and the rate of mulberry sac was the highest at 22 h maturation, and the effect of different maturation time on ICSI was observed in different maturation time of unwell sheep oocytes. The results showed that the cleavage rate of oocytes increased gradually with the increase of maturation time, and the mulberry sac rate of oocytes matured at 22 hours after maturation was the highest. The effect of hyperosmotic solution on ICSI was found to be the best time for ICSI. The results showed that hyperosmotic solution could significantly improve the successful rate of operation. But there was no significant difference in cleavage rate (43.9%) and mulberry sac rate (41.4%, 38.7%, P0.05). The effect of the first polar body position (6 and 12:00) on ICSI was not significant. The results showed that polar body position had no significant effect on ICSI cleavage rate (32.5 vs 34.5p 0.05) and mulberry sac rate (33.6% and 33.33.3% P0.05). (P 0.05) Ultrasonic treatment of different state sperm (fresh and frozen sperm) on ICSI. The results showed that ultrasonic treatment of fresh and frozen spermatozoa on oocyte fine oocyte. The cell cleavage rate (39.1 vs 37.4P0.05) and mulberry sac rate (33.7 vs 35.1and P0.05) had no significant difference (P 0.05).) the effects of different treatments of sperm (NaOH and ultrasound) on ICSI were not significant. The results showed that the effect of two treatments on oocyte cleavage rate 37.4 vs 46.6% (P0.05) the difference of mulberry sac rate in NaOH treatment group was significantly higher than that in ultrasonic treatment group (35.1g / L). The effect of co-incubation of spermatozoa on the oocyte development of ICSI was studied by using ICSI treated with NaOH as an example. There was no significant difference between the cleavage rate of ICSI embryos treated with NaOH and plasmids (37.9 vs 46.6% P0.05) and the rate of mulberry sac (45.3 vs 52.1% P0.05). The effect of non-co-culture and cumulus cell co-culture system on ICSI embryos was not significant. The results showed that the effects of co-culture and non-co-culture on ICSI embryos were not significant. There was no significant effect on cleavage rate 41.7% vs 44.0% P0.05) and mulberry sac rate 41.8% vs 35.7P0.05).
【學(xué)位授予單位】：河南農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：S826

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