共查询到19条相似文献，搜索用时 828 毫秒
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为了提高牛体外受精性控胚胎发育率,试验从体外受精（IVF）培养8 ｈ后的胚胎,采用不同的脱除卵丘细胞方式（机械脱除法、酶脱除法、振荡法）脱除卵丘细胞,研究影响牛性控精液体外受精效率的关键技术。结果表明,在体外受精8 h后,采用不同的脱除卵丘细胞方式（机械脱除法、酶脱除法、振荡法）脱除卵丘细胞,受精卵经体外培养48 h后统计胚胎卵裂率分别为（58.8±2.8）%、（58.5±5.7）%、（69.9±3.4）%,体外培养192 h统计囊胚率分别为（26.7±2.7）%、（26.9±3.9）%、（35.8±4.1）%,由此可见,振荡法显著优于机械脱除法和酶脱除法（P〈0.05）。性控精液体外受精时,体外受精8 h后采用振荡法脱除卵丘细胞对于提高性控精液体外受精胚胎发育率具有重要意义。 相似文献
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本文研究了淘汰奶牛卵巢采集方法、卵丘细胞和颗粒细胞对卵母细胞体外受精后发育的影响.结果表明奶牛屠宰后取其卵巢,用剖解法可比抽吸法得到更多的可用卵母细胞(8.5:6.2枚/卵巢,P＜0.05).经成熟培养和体外受精后,无卵丘细胞包围的裸露卵母细胞卵裂率为40.2%,但仅28.2%停留在8～16细胞期,最高发育阶段为16细胞期;而有3层以上卵丘细胞紧密包围的卵丘-卵母细胞复合体,其卵裂率(60.2%:53.2%,P＜0.05)和囊胚发育率(31.5%:19.6%,P＜0.05)均高于卵丘细胞包被不全的卵母细胞.在培养液中添加颗粒细胞,提高囊胚细胞数(96.2±5.2:70.4±4.6,P＜0.05),但没有促进牛受精卵的卵裂率. 相似文献
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通过比较参与受精的卵母细胞颗粒细胞存在与否、精子上浮时间、精卵共孵育时间、不同受精液等4个方面的因素,研究这些因素对猪卵母细胞体外受精后胚胎发育能力的影响,以求找到最佳的猪卵母细胞体外受精体系。将选择带有不同颗粒细胞的卵丘卵母细胞复合体分为3组：含全部颗粒细胞、2~3层颗粒细胞和裸卵;调整精子在受精液里的上浮时间为0、30、60、120min研究其受精能力;比较3、6、20h精卵共孵育时间对体外受精的影响;结果表明：在本试验体系下,在mTBM受精液中,将精子上浮处理60min,与含2~3层颗粒细胞的卵丘-卵母细胞复合体共孵育6h的IVF体系最为有效,其卵裂率为（77.6±2.3）%,囊胚率为（25.7±2.6）%。 相似文献
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利用树鼩卵母细胞的成熟培养、体外受精等方法,生产树鼩的试管婴儿,期望通过此路径来实现生产转基因树鼩动物模型。使用促性腺激素进行树鼩超数排卵、TCM199完全培基对卵母细胞进行体外成熟培养、卵母细胞与体外获能的附睾精子进行体外受精、受精卵发育至桑椹胚、囊胚的实验。结果表明,卵母细胞成熟率A级76.7%、B级55.01%、C级17.39%。受精率52%,受精卵采用体细胞共培养与非共培养的分裂率分别为37.14%和9.6%。桑椹胚/囊胚发育率分别为13.57%和0,(P<0.05)。树鼩卵母细胞的成熟培养、体外受精实验方法可行,受精卵在体外能发育到囊胚阶段。 相似文献
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试验利用水牛卵泡液(BuFF)和黄牛卵泡液(BoFF)对不同来源水牛卵母细胞体外受精效果的影响进行了探讨,以完善水牛体外受精培养系统,进一步提高水牛胚胎体外生产效率。试验按成熟培养液中添加卵泡液替代胎牛血清量共分4个组。不添加卵泡液(0%+10%胎牛血清)为Ⅰ组(对照组);添加5%卵泡液+5%胎牛血清为Ⅱ组;添加10%卵泡液+0%胎牛血清为Ⅲ组;添加15%卵泡液+0%胎牛血清为Ⅳ组。结果表明,添加BuFF对活体采集卵母细胞和屠宰场收集卵母细胞的体外受精卵分裂率无显著影响(P0.05),但添加5%和10%BuFF对卵母细胞体外受精后的胚胎发育有明显促进作用,囊胚率均极显著高于对照组和15%BuFF组(P0.01),5%和10%BuFF组间无显著差异(P0.05);添加15%BuFF囊胚率有降低的趋势,但与对照组相比差异不显著(P0.05)。而添加10%BoFF组活体采集卵母细胞体外受精的受精卵分裂率和囊胚率均极显著高于对照组和5%BoFF组(P0.01),添加5%BoFF组的受精卵分裂率和囊胚率与对照组无显著差异(P0.05);添加BoFF对屠宰场收集水牛卵母细胞体外受精卵分裂率无显著差异(P0.05),但添加5%和10%BoFF组的囊胚率均显著高于对照组和15%组(P0.05),添加15%BoFF组与对照组相比,囊胚率显著降低(P0.05),5%和10%BoFF组间囊胚率无显著差异(P0.05)。综合以上结果,在水牛卵母细胞成熟培养液中添加5%～10%的BuFF或BoFF代替牛血清,可明显提高水牛体外胚胎生产效率,且以添加同种的BuFF效果略好。 相似文献
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牦牛卵母细胞的体外成熟、种间受精与胚胎培养 总被引：1，自引：0，他引：1
探讨了卵母细胞体外成熟时间、卵母细胞质量、精子准备和受精卵培养体系对牦牛卵母细胞种间体外受精效果的影响。结果表明:牦牛卵母细胞随体外成熟培养时间延长,第一极体排出率增加,囊胚发育率以成熟培养24 h最高;A级卵母细胞种间受精后囊胚的发育率(48.77±3.76)%显著高于B级(32.05±5.24)%和C级(7.54±7.18)%(P<0.05);BO液洗涤离心处理的精子受精后卵裂率(82.53±6.54)%显著高于Percoll液分离精子受精后的卵裂率(67.39±4.50)%(P<0.05),而两者囊胚率差异不显著((42.32±4.13)%vs(35.59±5.62)%,P>0.05);荷斯坦奶牛精子浓度在1×106～5×106/mL范围与牦牛卵母细胞受精效果差异不显著。卵丘细胞、输卵管上皮细胞共培养和SOF液培养种间受精卵,其卵裂率差异不显著,但共培养组的桑葚胚、囊胚和孵化胚发育率均显著高于SOF液培养组。 相似文献
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卵丘细胞对卵母细胞成熟、受精和胚胎发育的影响试验 总被引：2，自引：0，他引：2
将从猪卵巢上获取的卵母细胞根据卵丘细胞层数的多少分为两类,并将这两类卵母细胞进行成熟培养、体外受精和孤雌激活,以验证两类卵母细胞的发育潜能.结果显示,将两类卵母细胞分别进行成熟培养时,优级卵的成熟率高于次级卵（67.42±177;1.52％vs 48.33±177;2.85％）,且差异极显著（P＜0.01）;将成熟的卵母细胞进行孤雌激活后,优级卵的囊胚率高于次级卵（58.00±177;2.88％ vs 41.00±177;6.40％）,且差异极显著（P＜0.01）,但囊胚总细胞教两者无显著差异（P＞0.05）;将成熟的卵母细胞进行体外受精后,优级卵的囊胚率、囊胚总细胞数均高于次级卵（18.00土5.70％ vs 4.25±177;2.31％,41.7±177;3.33 vs 36.2土2.10）,且差异极显著（P＜0.01）;进行受精卵孵育时,未去除卵丘细胞的胚胎卵裂率、囊胚率、囊胚总细胞数均高于去除卵丘细胞的（88.14土7.48％ vs 58.69±177;2.89％,51.2±177;7.33％ vs 11.92±177;2.29％,41.7±177;3.33 vs 36.8±177;3.15）,且差异极显著（P＜0.01）.以上结果说明：卵丘细胞层数较多的卵母细胞成熟率高,且其孤雌和体外受精胚胎的发育潜能也好;卵丘细胞存在时,有助于卵母细胞的受精及以后的胚胎发育. 相似文献
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Influence of co‐culture with denuded oocytes during in vitro maturation on fertilization and developmental competence of cumulus‐enclosed porcine oocytes in a defined system 
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下载免费PDF全文 Ruth Appeltant Tamás Somfai Kazuhiro Kikuchi Dominiek Maes Ann Van Soom 《Animal Science Journal》2016,87(4):503-510
Co‐culture of cumulus‐oocyte complexes (COCs) with denuded oocytes (DOs) during in vitro maturation (IVM) was reported to improve the developmental competence of oocytes via oocyte‐secreted factors in cattle. The aim of the present study was to investigate if addition of DOs during IVM can improve in vitro fertilization (IVF) and in vitro culture (IVC) results for oocytes in a defined in vitro production system in pigs. The maturation medium was porcine oocyte medium supplemented with gonadotropins, dbcAMP and β‐mercaptoethanol. Cumulus‐oocyte complexes were matured without DOs or with DOs in different ratios (9 COC, 9 COC+16 DO and 9 COC+36 DO). Consequently; oocytes were subjected to IVF as intact COCs or after denudation to examine if DO addition during IVM would affect cumulus or oocyte properties. After fertilization, penetration and normal fertilization rates of zygotes were not different between all tested groups irrespective of denudation before IVF. When zygotes were cultured for 6 days, no difference could be observed between all treatment groups in cleavage rate, blastocyst rate and cell number per blastocyst. In conclusion, irrespective of the ratio, co‐culture with DOs during IVM did not improve fertilization parameters and embryo development of cumulus‐enclosed porcine oocytes in a defined system. 相似文献
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添加不同发情时期山羊血清对牛胚胎体外发育的影响 总被引：1，自引：0，他引：1
为了提高牛体外胚胎囊胚发育率,添加不同发情时期山羊血清对牛胚胎进行体外培养。分别采集发情周期D0、D2、D4和D7山羊血清（发情当天为D0）,灭活除菌备用。结果表明不同发情时期羊血清对孤紫激活胚胎孵裂率影响差异不显著,囊胚发育率分别为29.5%、32.4%、33.9%和41.3%,D7血清能显著提高囊胚发育率（P＜0.05）,且扩张和孵化胚出现较早。添加D0血清体外受精胚胎卵裂率较高,但对囊胚发育率影响差异不显著。说明孤紫激活和体外受精胚胎发育有差异,D7血清对胚胎后期发育有利。受精胚无血清培养3d后添加D7血清,囊胚发育率为42.9%,是较为理想的培养方法。 相似文献
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Vascular endothelial growth factor (VEGF) promotes the early development of bovine embryo in the presence of cumulus cells 总被引：9，自引：0，他引：9
Luo H Kimura K Aoki M Hirako M 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2002,64(11):967-971
Three experiments were conducted to determine the effect of Vascular Endothelial Growth Factor (VEGF) on bovine embryonic development in vitro. Human recombinant VEGF(165) was employed at 5 ng/ml in modified synthetic oviduct fluid. In Exp. 1, bovine cumulus oocyte complexes were matured with or without VEGF for 22 hr, inseminated without VEGF for 6 hr, then cultured with or without VEGF for 42 hr. The cleavage rate and the development rate to 4- to 8-cell were higher (P<0.05) in groups with VEGF during in vitro maturation (IVM, 71.4% and 59.6%), in vitro culture (IVC, 70.3% and 62.3%), and both IVM and IVC (75.9% and 67.8%) than in the group cultured thoroughly without VEGF (49.9% and 38.4%, respectively). In Exp. 2, 4- to 8-cell embryos produced in vitro without VEGF were removed from cumulus cells at 48 hr post-insemination (Pi) and cultured with or without VEGF for 144 hr. The development rates to blastocyst at 96 hr (D6), 120 hr (D7) and 144 hr (D8) were similar (P>0.05) in both groups. In Exp. 3, cumulus cells were removed from presumptive embryos produced by IVM and IVF without VEGF at 10 hr Pi. Denuded embryos were cultured with or without VEGF for 38 hr or 182 hr. The cleavage rate and the development rates to 4- to 8-cell at 48 hr Pi and to blastocyst on D6, D7 and D8 were similar (P>0.05) in all groups. These results suggest that VEGF has a beneficial effect on the initial development of bovine embryo through surrounding cumulus cells. 相似文献
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Somfai T Kashiwazaki N Ozawa M Nakai M Maedomari N Noguchi J Kaneko H Nagai T Kikuchi K 《The Journal of reproduction and development》2008,54(3):149-155
The aim of the present study was to investigate the effects of centrifugation pretreatment on the viability and nuclear status of porcine in vitro matured (IVM) oocytes and on the developmental competence of in vitro fertilized (IVF) oocytes (zygotes) after cryopreservation by vitrification (Solid Surface Vitrification; SSV). Mature oocytes having the first polar body after IVM and zygotes having the second polar body at 10 h after IVF were centrifuged at 10,000 x g at 37 C for 20 min and then subjected to SSV. Their viability was evaluated by morphological appearance and fluorescein diacetate staining. The nuclear status of oocytes was evaluated 6 h after vitrification. The developmental ability to the blastocyst stage of vitrified zygotes was evaluated after 6 days of in vitro culture. Although centrifugation did not damage the oocytes directly, it drastically reduced the rate of live oocytes after SSV. The rates of vitrification-induced parthenogenetic activation were similar in both centrifuged and non-centrifuged oocytes (42.4 and 47.4%, respectively). Centrifugation had no significant effects on the viability of pronuclear oocytes. The development of vitrified zygotes to the blastocyst stage was significantly lower than that of the control irrespective of centrifugation pretreatment. There was no difference in the cleavage and blastocyst rates between the control and centrifuged zygotes after vitrification. There was also no difference in the total cell numbers of blastocysts between the control and centrifuged zygotes irrespective of vitrification. These results reveal that, in IVM porcine oocytes, centrifugation pretreatment is highly detrimental to cryotolerance; however, in zygotes, it has only a slight effect on viability and does not alter the developmental competence of surviving zygotes. 相似文献
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本研究利用卵母细胞体外成熟(IVM)、体外受精(IVF)技术,体外生产牛胚胎,以获得试管牛.结果表明:卵母细胞体外培养22～24h后,其成熟率为85％;体外受精率为83％.体外受精卵分别在颗粒细胞单层(GCM)和输卵管细胞单层(OCM)上培养,其胚胎最后产量(以授精时卵母细胞的数目为基数)分别为19％和29％,差异极显著(P<0.01).若体外受精卵先在GCM中培养72h后,再将已发生卵裂的(>4细胞)胚胎移到OCM中培养,其胚胎最后产量为35％.由此表明,早期胚胎在其发育过程中至少存在着3个发育相期,即1～8细胞、8～16细胞和16细胞～囊胚3个阶段,各阶段需要不同的培养环境.IVM/IVF胚胎移植到受体黄牛体内后,其足月时的妊娠率为15％.第一头试管犊牛已于1993年4月18日凌晨产出. 相似文献
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Effect of the Removal of Cumulus Cells on the Nuclear Maturation, Fertilization and Development of Porcine Oocytes 总被引：7，自引：0，他引：7
P Wongsrikeao Y Kaneshige R Ooki M Taniguchi B Agung M Nii T Otoi 《Reproduction in domestic animals》2005,40(2):166-170
The present study was conducted to investigate the effects of attachment of cumulus cells to porcine oocytes during the process of maturation and fertilization on the nuclear maturation, fertilization and subsequent development after in vitro fertilization (IVF). In the first experiment, the cumulus cells were removed from cumulus-oocyte complexes (COCs) at 0, 24 and 42 h after the onset of maturation culture and were then cultured until reaching 42 h of cultivation. In the second experiment, COCs were denuded as described in the first experiment, then fertilized and cultured for 7 days. As a control, cumulus cells were allowed to maintain attachment to the oocytes until the end of IVF. The proportion of oocytes reaching metaphase II significantly increased with the delay in the removal treatment of cumulus cells. The proportion of normal fertilization gradually increased with delay in the removal treatment of cumulus cells from COCs until the end of IVF. However, no significant difference in the proportion of normal fertilization was found between the 42-h and control groups. The removal treatment of cumulus cells in the 0- and 24-h group significantly (p < 0.05) decreased the proportion of cleaved embryos when compared with the control, and none of them developed to the blastocyst stage. The proportion of development to the blastocyst stage was significantly higher (p < 0.05) in the control group than in the 42-h group (18.1% vs 12.4%; p < 0.05). The present study indicates that the attachment of cumulus cells to the oocyte during maturation and fertilization is important to support oocyte nuclear maturation, fertilization and subsequent embryo development. Particularly, the attachment of cumulus cells to the oocyte during IVF promotes embryonic development. 相似文献
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Takada L Junior AM Mingoti GZ Balieiro JC Cipolla-Neto J Coelho LA 《Research in veterinary science》2012,92(1):124-127
The effect of melatonin during in vitro maturation (IVM) on DNA damage of cumulus cells (CCs) from bovine cumulus-oocyte complexes (COCs) and embryo development was evaluated. COCs from abattoir ovaries were cultured in maturation medium (MM) with 0.5μg/ml FSH and 5.0μg/ml LH (FSH-LH); 10(-9)M melatonin (MEL) or FSH-LH+MEL (FSH-LH-MEL). After 24h of in vitro maturation, the CCs surrounding the oocyte were subjected to DNA analysis by Comet assay. After in vitro fertilization and in vitro embryo culture, the embryo development rates were evaluated on day 2 post insemination (cleavage) and days 7-8 (blastocyst). The percentage of CCs with no DNA damage was significantly superior in MEL group (37.6±2.4) than in FSH-LH-MEL (28.0±2.4) and FSH-LH (17.8±2.41) groups. Cleavage and blastocysts rates were similar among groups. Melatonin during IVM protects the CCs from DNA damage but this effect did not influence embryo development in vitro. 相似文献