共查询到18条相似文献，搜索用时 421 毫秒
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①用EFS30、EFS40、EDFS30、EDFS40四种玻璃化冷冻液对MⅡ期水牛卵母细胞进行毒性试验,结果表明:试验组卵母细胞形态正常率与对照组均无显著性差异(P>0.05);对卵母细胞孤雌激活后EDFS30、EDFS40组的卵裂率与对照组(75.28%)及EFS30、EFS40组差异显著(P<0.05);利用4种冷冻保护剂采用OPS法冷冻保存MⅡ期水牛卵母细胞,其中以EDFS40作为冷冻液时,卵母细胞冷冻解冻后孤雌激活卵裂率最高,达31.60%;以EDFS40作为冷冻液,比较了GMP法和OPS法的冷冻效果,结果表明GMP法冷冻效果好于OPS法。②采用不同预处理时间和平衡时间使用细管法常规冷冻G V期卵母细胞,结果表明预处理5 min、平衡15min组的形态正常率和极体排出率相对较高,分别为72.73%、27.27%。 相似文献
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绵羊体外成熟卵母细胞OPS法玻璃化冷冻保存试验 总被引：1，自引：0，他引：1
研究以EDFS30为玻璃化冷冻液,以卵母细胞解冻后孤雌激活和体外受精后的卵裂率、囊胚发育率作为评价指标,探讨了以OPS法玻璃化冷冻保存体外成熟绵羊卵母细胞的效果。结果表明:卵母细胞孤雌激活后的卵裂率,冷冻组(64.2%)显著(P<0.05)低于毒性组(76.7%)和对照组(79.1%),而毒性组和对照组无显著(P>0.05)差异;卵母细胞孤雌激活后的囊胚发育率,冷冻组(4.2%)和毒性组(5.8%)均显著(P<0.05)低于对照组(20.2%),毒性组和冷冻组无显著(P>0.05)差异;冷冻组和毒性试验组卵母细胞体外受精后的卵裂率和囊胚发育率(67.6%和7.1%;62.3%和9.1%)均显著低于对照组(78.4%和28.4%)(P<0.05),而毒性组和冷冻组无显著(P>0.05)差异。可见以EDFS30为玻璃化冷冻液,采用OPS法冷冻保存绵羊体外成熟卵母细胞会在一定程度上降低其受精能力和胚胎发育能力。 相似文献
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本试验通过牛卵母细胞玻璃化冷冻过程中采用载体、冷冻时卵母细胞所带颗粒细胞层数和其成熟培养时间这3个方面对牛卵母细胞的玻璃化冷冻进行了探讨。(1)不同载体的对比:玻璃OPS管和塑料OPS管冷冻效果较好。玻璃OPS管冷冻卵母细胞,解冻后形态正常率、体外受精后卵裂率和囊胚率分别为91.2%、41.3%和14.52%。塑料OPS管冷冻卵母细胞,解冻后形态正常率、体外受精后卵裂率和囊胚率分别为89.09%、40.90%和14.54%。(2)保留2￣3层颗粒细胞的卵母细胞玻璃化冷冻的冷冻效果较好,其冷冻解冻后形态正常率、体外受精后卵裂率和囊胚率分别为90.79%、40.13%和15.13%。(3)体外成熟培养22h的卵母细胞冷冻效果较好,其冷冻解冻后形态正常率、体外受精后卵裂率和囊胚率分别为94.14%、41.80%和15.23%。 相似文献
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试验以屠宰场云岭黑山羊卵巢卵母细胞为材料,研究其玻璃化冷冻的效果。试验中选用20% EG+20% DMSO为冷冻液、冷冻环为载体,以20 s、40 s玻璃化时间冷冻GV和MⅡ期的卵母细胞。结果表明,GV期卵母细胞的形态正常率、成熟率和卵裂率都很低,且解冻成熟培养后冷冻组的成熟率和卵裂率极显著低于对照组(P<0.01)。而MⅡ期卵母细胞冷冻效果较好,毒性试验组和冷冻组形态正常率分别为91.1%和83.3%,明显高于GV期;孤雌激活后毒性组卵裂率与对照组无显著性差异(P>0.05),冷冻组的卵裂率显著低于对照组(P<0.05)。用20 s、40 s玻璃化时间冷冻的卵母细胞解冻后GV和MⅡ期各组均无显著差异。根据试验结果得出在冷冻保存中最好冷冻MⅡ期的卵母细胞,以便提高后期的卵裂率和囊胚率;卵母细胞玻璃化时间在40 s内均不影响卵母细胞的活力和发育潜力。 相似文献
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试验采用EFS30、DFS30和EDFS30冷冻液对小鼠MⅡ期卵母细胞进行玻璃化冷冻,并对冷冻-解冻后卵母细胞存活率、纺锤体形态正常率、孤雌激活胚胎的发育能力进行研究。结果表明:利用EDFS30冷冻-解冻卵母细胞后的存活率(98.3%)显著高于EFS30组(88.2%)和DFS30组(89.5%)(P0.05);DFS30组纺锤体形态正常率(44.9%)显著低于其他各组;而孤雌激活后EFS30、DFS30、EDFS30和对照组之间卵裂率(59.1%、56.3%、59.3%和60.0%)、囊胚率(52.0%、53.7%、61.2%和62.1%)和囊胚细胞数(49、48、50、50)均无显著性差异(P0.05)。综上所述,3种冷冻液冷冻卵母细胞孤雌激活后均可获得较好的体外发育能力,其中EDFS30较适宜小鼠MⅡ期卵母细胞冷冻保存。 相似文献
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试验采用3种冷冻载体(麦管、开放式拉长麦管和半麦管)对猪卵母细胞玻璃化冷冻后发育能力的影响进行研究,以探讨采用自制半麦管作为载体对猪MII期卵母细胞玻璃化冷冻后发育能力的影响.结果发现,采用半麦管法、OPS法玻璃化冷冻的猪卵母细胞的形态完整率、存活率、卵裂率分别为90.85%、60.07%、20.43%和84.15%、55.71%、14.71%,无显著差异(P＞0.05),但采用半麦管法的结果好于OPS法;与麦管法的形态完整率、存活率和卵裂率(分别为60.06%,39.64%和0)均有显著性差异(＜0.05).结论为采用半麦管法玻璃化冷冻猪卵母细胞可以稍微提高其冻后发育能力. 相似文献
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实验研究了不同成熟培养时间的牛卵母细胞玻璃化冷冻及胞质内单精子注射(ICSI)后的受精效果。结果表明:成熟后的新鲜牛卵母细胞按照ICSI注射方法穿刺而不注射精子组与未经穿刺的对照组相比,孤雌激活后的卵裂率、囊胚发育率及囊胚细胞数无显著差异(P>0.05);成熟培养16h(MⅠ)和23h(MⅡ)卵母细胞冷冻解冻后形态正常率均显著低于新鲜对照组(76.66%、87.33%vs100.0%)(P<0.05),冷冻解冻后二者分别成熟培养至24h,ICSI后胚胎的囊胚发育率(5.29%、14.41%)显著低于新鲜对照组(24.40%)(P<0.05);成熟培养23h与成熟培养16h的卵母细胞冷冻解冻后形态正常率及ICSI后囊胚发育率(14.41%vs5.29%)均有显著性差异(P<0.05)。实验证明,ICSI操作不会影响卵母细胞发育潜力;玻璃化冷冻影响卵母细胞解冻后形态正常率以及ICSI后胚胎的发育能力;成熟培养23h比16h的卵母细胞冷冻保存后经ICSI的胚胎发育潜力高。 相似文献
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玻璃化冻存对驴卵母细胞超微结构的影响 总被引：1，自引：1，他引：0
试验旨在探究玻璃化冷冻对驴卵母细胞发育的影响,寻求驴卵母细胞冷冻的最佳条件。通过对不同发育时期的驴卵母细胞进行玻璃化冷冻,冷冻复苏后分别进行成熟培养和孤雌激活,并对GV期未冷冻组(对照组)、GV期冷冻组、IVM-MⅡ冷冻组卵母细胞微丝和线粒体超微结构进行免疫荧光标记,统计冷冻复苏后卵母细胞形态正常率、成熟率、孤雌激活卵裂率、超微结构正常率。结果表明,GV期冷冻组卵母细胞的形态正常率与GV期未冷冻组(对照组)间无显著差异(P0.05),成熟率和卵裂率均显著低于对照组(P0.05);IVM-MⅡ冷冻组的卵裂率显著低于对照组(P0.05),且卵裂后细胞发育受到阻滞。冷冻组微丝在皮质区分布明显减少的卵母细胞数目增多,冷冻组卵母细胞的线粒体数量明显低于对照组,由此可以说明冷冻对卵母细胞超微结构有损伤,从而导致复苏后成熟率下降,影响卵母细胞的受精和体外发育,且GV期冷冻组较IVM-MⅡ冷冻组在微丝与线粒体结构上有较小损伤,发育状态较好。 相似文献
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GMP玻璃化冷冻水牛GV期和MⅡ期卵母细胞 总被引：1，自引：0，他引：1
采用玻璃微细管法(Glass micropipette vitrification,GMP)冷冻了水牛GV期和MⅡ期卵母细胞。结果显示,GV期冻后卵母细胞的形态正常率,孤雌激活后的发育潜力均明显低于未冷冻组(P<0.05)。在进一步探讨不同发育阶段的水牛卵母细胞冷冻保存效果中发现,GV期卵母细胞的形态正常率,体外成熟后的孤雌激活分裂率和8-细胞率显著低于MⅡ期(P<0.05)。结果表明,水牛GV期卵母细胞冷冻的效果比MⅡ期差。 相似文献
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水牛卵母细胞玻璃化冷冻保存 总被引：1，自引：0，他引：1
以水牛MII期的卵母细胞为材料,利用玻璃化冷冻液EDS33(16.5%EG+16.5%DMSO+sucrose)对水牛MII期的卵母细胞进行两步法(玻璃毛细管(GMP)和拉细的开口塑料细管(OPS))玻璃化冷冻保存,即卵母细胞首先放入预平衡液(7.5%EG+7.5%DMSO+sucrose)中平衡3 min,再移入玻璃化冷冻液中30 s后装管直接投入液氮.解冻是在蔗糖浓度逐渐降低的解冻液中进行的.解冻后存活的卵母细胞孤雌激活,通过囊胚发育率作为评定卵母细胞冷冻效果的指标.结果发现,GMP和OPS法冷冻保存的水牛卵母细胞解冻后的存活率(分别为96.80%和97.41%)与对照组卵母细胞的存活率(100%)3者之间差异均不显著(P>0.05).GMP法和OPS法冷冻的水牛卵母细胞激活后的胚胎分裂率和囊胚发育率2者均明显低于对照组(分别为30.58%和28.32% vs50.94%,10.81%和9.38% vs 29.63%,P<0.05),而这2种方法冷冻的水牛卵母细胞激活后的分裂率和囊胚发育率差异均不显著(P>0.05).这表明GMP和OPS玻璃化冷冻方法可以用于水牛卵母细胞的冷冻,并且玻璃化冷冻的卵母细胞能继续分裂并发育到囊胚. 相似文献
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KGhM Mahmoud TH Scholkamy YF Ahmed GE Seidel Jr MF Nawito 《Reproduction in domestic animals》2010,45(4):565-571
This study was designed to evaluate effects of different combinations of cryoprotectants on the ability of vitrified immature buffalo oocytes to undergo in vitro maturation. Straw and open‐pulled straw (OPS) methods for vitrification of oocytes at the germinal vesicle stage also were compared. The immature oocytes were harvested from ovaries of slaughtered animals and were divided into three groups: (i) untreated (control); (ii) exposed to cryoprotectant agents (CPAs); or (iii) cryopreserved by straw and OPS vitrification methods. The vitrification solution (VS) consisted of 6 m ethylene glycol (EG) as the standard, control vitrification treatment, and this was compared with 3 m EG + 3 m dimethyl sulfoxide (DMSO), 3 m EG + 3 m glycerol, and 3 m DMSO + 3 m glycerol. Cryoprotectants were added in two steps, with the first step concentration half that of the second (and final) step concentration. After warming, oocyte samples were matured by standard methods and then fixed and stained for nuclear evaluation. Rates of MII oocytes exposed to CPAs without vitrification were lower (54.3 ± 1.9% in EG, 47.5 ± 3.4% in EG + DMSO, 36.8 ± 1.2% in EG + glycerol and 29.9 ± 1.0% in DMSO + glycerol; p < 0.05) than for the control group (79.8 ± 1.3%). For all treatments in each vitrification experiment, results were nearly identical for straws and OPS, so all results presented are the average of these two containers. The percentages of oocytes reaching telophase‐I or metaphase‐II stages were lower in oocytes cryopreserved using all treatments when compared with control. However, among the vitrified oocytes, the highest maturation rate was seen in oocytes vitrified in EG + DMSO (41.5 ± 0.6%). Oocytes cryopreserved in all groups with glycerol had an overall low maturation rate 19.0 ± 0.6% for EG + glycerol and 17.0 ± 1.1% for DMSO + glycerol. We conclude that the function of oocytes was severely affected by both vitrification and exposure to cryoprotectants without vitrification; the best combination of cryoprotectants was EG + DMSO for vitrification of immature buffalo oocytes using either straw or OPS methods. 相似文献
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The breeding of Mangalica, a native pig breed in Hungary, had been started in 1833, but this pig breed almost became extinct in Hungary in the past decades. In 1991, the number of sows was only 200. Although in these days the existing Mangalica population consists of more than 6000 animals representing different colour variations, the preservation of this traditional pig breed is still very important. Vitrification is a potential tool for the preservation of gametes and embryos of these animals. The aim of this study was to investigate the effects of vitrification on the developmental competence of Mangalica (M) and Large White (LW) oocytes following fertilisation. The oocytes were vitrified by the Open Pulled Straw (OPS) method using different concentrations of ethylene glycol and dimethyl sulphoxide as cryoprotectants. After rehydration the oocytes underwent in vitro fertilisation; the resultant zygotes were then cultured in vitro for four days to assess embryonic development. In the first experiment, in vitro maturation of M and LW oocytes was compared. No significant difference was observed in the nuclear maturation rate of LW and M oocytes. In the second experiment, the sensitivity of oocytes to vitrification was examined by evaluating oocyte morphology after thawing. A higher percentage of LW oocytes showed normal morphology compared to M oocytes, indicating that Mangalica oocytes are more sensitive to cryoprotectants than Large White oocytes. After warming and in vitro fertilisation, more than 50% of the oocytes started embryonic development and by the end of the incubation period morula stage embryos had developed in both groups. The results show that the OPS vitrification technique is well suited to preserve Mangalica oocytes and from these oocytes morula embryos can be produced. 相似文献
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卵母细胞冷冻保存具有广泛而潜在的应用价值。文章主要分析了不同冷冻方法对绵羊GV期卵母细胞发育效果的影响。GV期卵母细胞程序化冷冻解冻后形态正常率（54.8%）和体外培养成熟率（14.7%）均显著或极显著低于细管玻璃化冷冻（71.8%,29.5%;P〈0.05）和OPS玻璃化冷冻（78.3%,P〈0.01;35.4%,P〈0.05）;3种不同方法冷冻GV期卵母细胞成熟率与对照组相比均差异极显著（P〈0.01）。 相似文献
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Kohaya N Fujiwara K Ito J Kashiwazaki N 《The Journal of reproduction and development》2011,57(6):675-680
Unfertilized oocytes are one of the most desired germ cell stages for cryopreservation because these cryopreserved oocytes can be used for assisted reproductive technologies, including in vitro fertilization (IVF) and intracytoplasmic sperm injection. However, in general, the fertility and developmental ability of cryopreserved oocytes are still low. The aim of the present study was to improve vitrification of mouse oocytes. First, the effects of calcium and cryoprotectants, dimethyl sulfoxide and ethylene glycol (EG), in vitrification medium on survival and developmental ability of vitrified oocytes were evaluated. Oocytes were vitrified by a minimal volume cooling procedure using different cryoprotectants. Most of the vitrified oocytes were morphologically normal after warming, but their fertility and development were low independently of calcium and cryoprotectants. Second, the effect of cumulus cells on ability of oocytes to be fertilized and develop in vitro was examined. The fertility and developmental ability of denuded oocytes (DOs) after IVF were reduced compared with cumulus-oocyte complexes (COCs) both in fresh and cryopreserved groups. Vitrified COCs showed significantly (P<0.05) higher fertility and ability to develop to the 2-cell and blastocyst stages than those of vitrified DOs with cumulus cells and vitrified DOs alone. The vitrified COCs developed to term at a high success rate equivalent to the rate obtained with IVF using fresh COCs. Taken together, the current results clearly demonstrate that, in the presence of surrounding cumulus cells, matured mouse oocytes vitrified using calcium-free media and EG retain their developmental competence. These findings will contribute to improve oocyte vitrification in not only experimental animals but also clinical application for human infertility. 相似文献
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二甲基亚砜(DMSO)、丙二醇(PROH)、乙二醇(EG)和甘油(GL)4种冷冻保护剂程序化冷冻牛GV期卵母细胞的结果表明,EG和PROH的保护效果比GL和DMSO好。4种不同冷冻方法冷冻保存牛GV期卵母细胞,比较解冻后卵母细胞的体外成熟率、受精后卵裂率。结果表明,在程序化冷冻法与细管玻璃化法(Straw)之间的差异不显著(P>0.05),在开放式拉管法(OPS)与毛细玻管法(GMP)之间的差异不显著(P>0.05);但OPS和GMP与程序化冷冻法和Straw之间的差异极显著(P<0.01)。玻璃化冷冻效果优于程序化冷冻。说明GMP和OPS玻璃化冷冻优于Straw玻璃化冷冻。说明可以采用GMP方法冷冻保存牛GV期卵母细胞。 相似文献
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采用PBS作为冷冻基础液,分别用甘油和二甲基亚砜(DMSO)作为冷冻保护液,在程序化冷冻保存和玻璃化冷冻保存条件下,研究小鼠生发泡期(GV期)卵母细胞的抗冻能力。结果表明,2种冷冻方法对小鼠GV期卵母细胞解冻后形态正常率和存活率无显著影响(P>0.05)。冷冻保护剂种类对小鼠GV期卵母细胞解冻后形态正常率无显著影响(P>0.05);但对存活率有显著影响,玻璃化冷冻采用二甲基亚砜作为冷冻保护液效果极显著优于甘油(P<0.01)。以冷冻效果较好的二甲基亚砜作为冷冻保护液,采用玻璃化冷冻不同发育阶段(GV期和MⅡ期)的小鼠卵母细胞,解冻后形态正常率无显著差异(P>0.05),但存活率GV期要显著优于MⅡ期卵母细胞(P<0.05)。 相似文献