共查询到18条相似文献，搜索用时 156 毫秒
1.
为探讨胰岛素(Insulin)和白血病抑制因子(Leukemia inhibit factor,LIF)对猪卵母细胞体外成熟(IVM)和猪孤雌激活胚胎(PAEs)的影响,在卵母细胞体外成熟或者胚胎培养基中添加Insulin和LIF,研究卵裂率和囊胚率的变化。结果:添加了5μg/mL Insulin后猪卵母细胞体外成熟效果显著提高,但成熟后孤雌激活发育能力与非添加组相近;而胚胎培养基中添加Insulin对孤雌胚的卵裂和囊胚的形成也没有明显促进作用;添加1 000 U/mL的LIF后,卵母细胞核成熟率没有明显提高,反而孤雌激活后囊胚率急剧下降,但对卵裂率以及囊胚总细胞数影响不大;在胚胎培养基中添加LIF后,孤雌胚的卵裂和囊胚形成并没有明显的提高。表明:Insulin对卵母细胞体外成熟有益,但是对孤雌胚胎的最佳处理程序还需要摸索;本文所采用的LIF处理对猪卵体外成熟以及孤雌胚胎体外发育没有帮助,还需要进一步研究其他浓度和处理程序对猪卵母细胞体外成熟和孤雌激活胚胎发育能力的影响。 相似文献
2.
Ghrelin对水牛体外受精和孤雌激活胚胎体外发育的影响 总被引：1，自引：0，他引：1
本研究的目的是探讨Ghrelin对水牛体外受精和孤雌激活胚胎体外发育的影响.体外成熟的水牛卵母细胞经体外受精或离子霉素孤雌激活后.分别在舍0,0.5,5,50和500 μg/L Ghrelin的培养液中进行体外培养,观察各组胚胎的卵裂率和囊胚率.结果显示,在培养液中添加不同浓度的Ghrelin对体外受精和孤雌激活胚胎的卵裂率均无显著影响(P>0.05),但添加500 μg/L的Ghrelin显著提高体外受精胚胎的囊胚发育率(33.5% vs 13.7%,P<0.05),50 μg/L或500 μg/L的Ghrelin均显著提高孤雌激活胚胎的囊胚发育率(32.4%和34.6% vs 14.5%,P<0.05).结果表明,培养液中添加Ghrelin对胚胎的早期卵裂没有影响,但可促进水牛体外受精和孤雌激活胚胎囊胚的形成. 相似文献
3.
4.
为了探讨瘦素(Leptin)对猪卵母细胞体外成熟及孤雌激活后胚胎早期发育的影响,研究选择在Earle's盐缓冲的TCM199中添加10IU/mLeCG,10IU/mLhCG,10ng/mLEGF配制成化学限定的基础液,以添加不同浓度Leptin设定各试验组,对猪卵母细胞进行体外成熟培养。以未添加Leptin的基础液为对照组1,而添加5%的胎牛血清(FBS)、10%猪卵泡液为对照组2。比较分析各组卵母细胞核成熟效率,孤雌激活后胚胎的卵裂率,囊胚发育率。结果表明:各添加组卵母细胞成熟率与对照组之间无显著差异(P>0.05);孤雌激活后,各组间的卵裂率和囊胚发育率也无明显差异。同时,在化学限定的猪卵母细胞体外成熟液中添加Leptin对猪卵母细胞体外成熟和孤雌激活后早期胚胎的发育无显著效果。 相似文献
5.
《中国兽医学报》2015,(9)
旨在探索宁夏滩羊卵母细胞孤雌激活及胚胎培养条件,并建立转Cherry基因滩羊体细胞重构胚的融合与体外培养体系。试验分析了3种激活方法即电激活、Ionomycin结合6-DMAP与电激活结合6-DMAP对成熟的滩羊卵母细胞激活的影响,以及3种胚胎培养液mSOFaa、M16和KSOM的培养筛选,并优化了滩羊体细胞重构胚的融合与体外培养条件。结果表明:在卵母细胞孤雌激活中,最适电压为1 600V/cm,获得卵裂率和囊胚率分别为58.5%和19.5%;5μmol离子霉素结合2 mmol 6-DMAP能有效地激活成熟的滩羊卵母细胞,卵裂率为81.0%,囊胚率为26.2%;并发现mSOFaa胚胎培养液的卵裂率和囊胚显著优于M16和KSOM培养液;在转基因重构胚融合中发现在电压1 800V/cm、脉冲时间10μs、脉冲次数2次和间隔时间为1s的条件下,卵裂率和囊胚率为40.0%、21.4%。本试验建立的滩羊孤雌激活和转基因重构胚融合与体外培养体系为宁夏滩羊的分子育种奠定了基础。 相似文献
6.
《中国兽医学报》2014,(6):995-998
在NCSU-23改良培养液中分别添加不同质量浓度的脑源性神经生长因子(不rain-derived neurotrophic factor,BDNF)和神经生长因子(nerve growth factor,NGF),观察其对猪孤雌激活胚胎体外生长的影响。结果显示:在培养液中添加40μg/L的BDNF,可促进猪孤雌激活胚胎的囊胚形成;在猪PA胚胎培养液中添加不同质量浓度的NGF,对猪孤雌激活胚胎的在卵裂率、囊胚率和囊胚孵化率影响不明显。结果表明,在NCSU-23改良培养液中添加一定质量浓度的BDNF可促进猪孤雌激活胚胎体外培养过程中囊胚的形成。 相似文献
7.
体外成熟对牛卵母细胞孤雌激活后发育潜力的影响 总被引：2，自引：1，他引：1
本试验在卵母细胞体外成熟的研究基础上,研究了培养用水、卵泡液和成熟时间对卵母细胞体外成熟及孤雌激活后发育潜力的影响.结果表明(1)将卵母细胞分别于蒸馏水和Milli-Q超纯水配制的成熟培养液中培养22～24 h, 孤雌激活后, 卵裂率无显著差异(P>0.05); 但孤雌卵在超纯水配制的胚胎培养液中培养,囊胚发育率为22.3%, 明显高于在蒸馏水配制的培养液中的囊胚发育率14.1%(P<0.05).(2)在成熟培养液中添加10%、20%卵泡液,成熟卵母细胞孤雌激活后的囊胚发育率(32.3%, 30.9%)明显高于添加5%和0%卵泡液组的囊胚发育率(21.8%, 22.7%,P<0.05).卵母细胞在添加20%卵泡液的培养液中成熟培养,孤雌激活后囊胚孵化率明显低于添加0、5%、10%卵泡液组的囊胚孵化率(P<0.05).(3)成熟28 h或30 h的卵母细胞孤雌激活后,其囊胚发育率(30.5%或29.4%)明显高于成熟24 h或26 h组的囊胚发育率(20.5%或22.1%,P<0.05). 相似文献
8.
为探讨表皮生长因子(epidermal growth factor,EGF)的添加浓度及脱卵丘细胞时间对猪卵母细胞体外成熟及孤雌胚胎体外发育的影响.试验通过在体外成熟液中添加不同浓度(0、10、15、20、30、40 ng/mL)的EGF来研究其对培养44 h的卵母细胞成熟率以及孤雌胚胎发育的影响;在培养开始后的不同时间(18、24、38、44 h)进行脱卵丘细胞处理来研究不同时间脱卵丘处理对培养44 h的卵母细胞成熟率以及孤雌胚胎发育的影响.结果表明,成熟培养基中添加10 ng/mL EGF能显著提高卵母细胞的卵裂率和囊胚率(P <0.05).共培组和独培组卵母细胞培养18 h后脱卵丘细胞成熟率均低于44 h,但差异不显著(P >0.05);共培组卵母细胞培养18 h后脱卵丘细胞的卵裂率和囊胚率显著高于培养44 h(P <0.05);独培组卵母细胞培养18 h后脱卵丘细胞的卵裂率与44 h无显著差异(P >0.05),但囊胚率显著高于培养44 h后脱卵丘细胞(P <0.05).添加10 ng/mL EGF对猪卵母细胞体外成熟及孤雌胚胎体外发育较好;卵母细胞培养18 h后脱卵丘细胞可提高孤雌胚胎早期发育能力. 相似文献
9.
为了研究曲古抑菌素A(TSA)和5-氮杂-2’-脱氧胞苷(5-Aza-CdR)对猪孤雌胚胎发育及胚胎质量的影响,试验采用猪卵母细胞孤雌激活的方法,孤雌激活后在胚胎培养液中分别添加不同浓度TSA和5-Aza-CdR,比较其对猪孤雌胚胎发育的影响。结果表明:40 nmol/L TSA处理24 h能显著提高孤雌胚胎的囊胚率及囊胚细胞个数(P<0.05),卵裂率无明显变化(P>0.05);30 nmol/L5-Aza-CdR处理48 h能显著提高孤雌胚胎的囊胚率(P<0.05),卵裂率与囊胚细胞个数均无明显变化(P>0.05)。说明在一定浓度条件下,TSA和5-Aza-CdR对猪孤雌胚胎发育的囊胚率有显著促进作用,5-Aza-CdR处理对卵裂率、囊胚细胞个数的影响不大,但TSA处理可以明显提高囊胚细胞个数,从而提高胚胎质量。 相似文献
10.
研究旨在探讨单宁酸对猪卵母细胞体外成熟质量及其胚胎发育能力的影响。在猪卵丘卵母细胞复合体（COCs）体外成熟培养液中添加不同浓度（0、1、10、100 μg/mL）单宁酸培养42 h后,检测COCs的扩散程度和卵丘细胞扩散指数,统计COCs的体外成熟率,检测成熟卵母细胞内谷胱甘肽（glutathione,GSH）、活性氧（reactive oxygen species,ROS）和生长分化因子9（growth differentiation factor 9,GDF9）的水平,并统计孤雌激活及体外受精胚胎48和168 h的卵裂率、囊胚率及囊胚总细胞数。结果显示,与对照组相比,10 μg/mL单宁酸组卵丘细胞扩散指数显著提高（P<0.05）,100 μg/mL单宁酸组显著降低（P<0.05）;1和10 μg/mL单宁酸组卵母细胞成熟率差异不显著（P>0.05）,100 μg/mL单宁酸组卵母细胞成熟率显著降低（P<0.05）;1和10 μg/mL单宁酸组GSH和GDF9水平显著提高（P<0.05）,ROS水平显著降低（P<0.05）。孤雌胚胎和体外受精胚胎发育能力结果显示,与对照组相比,各单宁酸组卵裂率差异不显著（P>0.05）,10 μg/mL单宁酸组孤雌胚胎囊胚率及体外受精胚胎囊胚率显著提高（P<0.05）,100 μg/mL单宁酸组孤雌胚胎囊胚细胞数及体外受精胚胎囊胚细胞数均显著低于其他各组（P<0.05）。以上结果表明,10 μg/mL单宁酸可通过提高卵丘细胞扩散能力及GSH和GDF9水平、降低卵母细胞内ROS水平,改善猪卵母细胞成熟质量,提高孤雌胚胎及体外受精胚胎的发育能力。 相似文献
11.
探讨电激液中不同Ca2+浓度与不同电脉冲强度相互关系对猪体外成熟卵母细胞孤雌激活的影响;以及山梨醇电激活液在猪孤雌激活中的应用。结果表明,当Ca2+浓度不高时,同时增加电激液Ca2+浓度和电脉冲强度能协同促进孤雌胚胎的卵裂率和囊胚率的提高,Ca2+浓度为0.05和0.1mmol/L时,分别以1.6kV/cm和1.2kV/cm激活,得到的卵裂率为73.75%、74.70%;囊胚率为37.50%、36.83%,显著高于其余各组(P0.05);当Ca2+过高,增加脉冲强度卵母细胞孤雌发育能力反而下降,退化率升高;以山梨醇液进行电激活,在1.6kV/cm时,卵裂率和囊胚率最高,分别为77.23%和34.15%;与甘露醇电激液不同,施加交流电,山梨醇液不能对孤雌胚胎发育起到促进作用;将山梨醇、甘露醇电激液等体积混合,交流脉冲后进行电激活,1.2、1.6kV/cm组卵裂率分别为72.33%和70.03%,囊胚率分别为31.03%和29.60%,虽然略低于甘露醇组(77.07%、36.03%),但优于山梨醇组(69.63%、26.93%),差异不显著(P0.05)。以上结果说明,猪卵母细胞激活所需内流Ca2+浓度存在临界值,临界值内,提高Ca2+浓度或电参数,都能提高卵母细胞的激活效果;超过临界值,效果相反;山梨醇液能够取代甘露醇液用于猪卵细胞的电激活;同时以山梨醇和甘露醇作为电激液的基本成份,完全可以用于猪卵母细胞的电激活或融合。 相似文献
12.
Y Tao H Chen NN Tian DT Huo G Li YH Zhang Y Liu FG Fang JP Ding XR Zhang 《Reproduction in domestic animals》2010,45(1):19-25
The maturation and developmental potential on cumulus-cell-free oocytes is of great importance theoretically and practically. The present study was to investigate the effects of l -ascorbic acid, α-tocopherol and co-culture on in vitro developmental potential of porcine denuded oocytes (DOs). Porcine DOs were cultured in maturation medium supplemented with vitamin C (0, 50, 100, 250, 500, 750 μM) and vitamin E (0, 10, 20, 50, 100, 250 μ m ), respectively. And they were also co-cultured with dispersed cumulus cells (group CCscoculture), intact cumulus cells oocyte complexes (COCs) (group COCscoculture), and COCs whose oocytes were removed (group OOXcoculture), respectively. After 44 h incubation, the maturation rates, cleavage rates and blastocyst rates after parthenogenetic activation in three experiments mentioned above were collected and analysed, respectively. L -Ascorbic acid promoted porcine DOs in vitro maturation and blastocyt development after parthenogenetic activation while α-tocopherol did not increase the in vitro maturation rates, but improved the blastocyst rate. None of the three co-culture manner promoted the in vitro maturation and the cleavage of porcine DOs after parthenogenetic activation, but all the co-culture manners improved the blastocyst rates. Both Vitamin C and E enhance the in vitro developmental potential of porcine DOs. Co-culture increases the developmental potential of porcine DOs. 相似文献
13.
Karja NW Otoi T Murakami M Wongsrikeao P Budiyanto A Fahrudin M Nagai T 《The Journal of reproduction and development》2005,51(6):783-786
This study was conducted to improve parthenogenetic development in vitro of feline oocytes following a combined activation treatment of electrical stimulation and cycloheximide. In vitro matured (IVM) oocytes were stimulated electrically by a DC electrical pulse of 2 kV/cm for 50 micros. The stimulated oocytes were then incubated in MK-1 medium with or without cycloheximide and subsequently cultured in vitro for 6 days. No significant differences were observed between the two groups with respect to the proportions of cleavage, development to the morula stage, and the cell number of blastocysts. However, exposure of electrically stimulated oocytes to cycloheximide significantly increased the rate of development of the stimulated oocytes into the blastocyst stage compared with oocytes stimulated by electrical stimulation alone (31.0% vs 6.7%). The results from the present study suggested that a single electrical stimulation was insufficient to activate the IVM cat oocytes at 24 h of maturation and that exposure to cycloheximide following electrical stimulation improved the efficacy of the parthenogenetic development of domestic cat oocytes. 相似文献
14.
Bing Y Che L Hirao Y Takenouchi N Rodríguez-Martínez H Nagai T 《The Journal of reproduction and development》2003,49(2):159-166
This study was designed to evaluate the parthenogenetic activation of porcine oocytes matured in vitro for a varied period after combined electric pulse (EP; 1500 V/cm, 100 microsec) and Butyrolactone I (BL I). After 36 h of maturation culture, the rates of activated oocytes and oocytes with two pronuclei were significantly lower than those of oocytes cultured for 42 and 48 h after EP. However, when treated by a combined EP and BL I (150 microM), these rates increased to the same level as 42 and 48 h oocytes. When oocytes cultured for 48 h and activated by a combined EP and BL I treatment were subsequently cultured in mNCSU37 medium, the rates of embryos cleaved and developed to the blastocyst stage were significantly higher than those in Whitten's medium. In contrast, when activated oocytes were cultured in mNCSU37 medium under two oxygen environments (5% vs 20% O(2)), there was no difference in the rates of cleavage, blastocyst formation and nuclear numbers per blastocyst. Our results demonstrated that the combined EP and BL I treatment of porcine oocytes matured in vitro is capable of producing high rates of good quality blastocysts when cultured in a suitable in vitro condition. 相似文献
15.
Hyun S Lee B Lee G Lee E Lim J Kang S Hwang W 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2003,65(1):51-56
To evaluate whether oocytes excluded from somatic cell nuclear transfer (SCNT) could be utilized for embryo production by parthenogenetic activation (PA), porcine oocytes with poor morphology after maturation culture were excluded from SCNT and subsequently used for PA with different stimuli. In the first set of experiment, either electric pulse of different strengths (1.75, 2.0 or 2.25 kV/cm for 30 microsec each) or chemicals with different treatment durations [7% ethanol for 5 min followed by exposure to 6-dimethylaminopurine (6-DMAP) for 0, 2, 3 or 4 hr] was employed. Development to the 8-cell and morula stages was significantly (P<0.05) improved by electric stimulation of 2.0 kV/cm, while blastocyst formation was enhanced by chemical treatment of ethanol and 6-DMAP for 4 hr. Subsequently, oocytes were parthenogenetically activated by one of four stimuli; 1) optimal electric (2.0 kV/cm for 30 microsec), 2) optimal chemical (ethanol followed by 6-DMAP for 4 hr), 3) electric then chemical and 4) vice versa. On the other hand, oocytes with normal morphology were subjected to the same experimental treatments for the control. Regardless of oocyte type, a combination of electric and chemical stimulations did not further stimulate preimplantation development, compared with electric activation only. However, combinational treatment greatly increased the cell number of blastocysts in SCNT-excluded oocytes (21.9 to 22.9 vs. 16.9 cells/blastocyst), while such effect was not found in normal oocytes (22.2 to 23.3 cells/blastocyst). In conclusion, porcine oocytes excluded from SCNT still have a potential to develop blastocysts after PA and this might contribute to increasing the efficiency of SCNT for various purposes. A combined activation by electricity and chemical yielded the best rate of preimplantation development with increasing the quality of blastocyst. 相似文献
16.
采用NCSU-37为体外成熟、受精、培养体系,比较不同成熟时间46h、58h和70h的猪卵母细胞对体外受精的影响和孤雌发育。结果显示,猪卵母细胞在体外成熟培养46h、58h和70h后,46h培养组的卵母细胞体外受精后的卵裂率明显低于58h和70h培养组(P<0.05),但囊胚发育率明显高于其他两组(P<0.05)。46h组的孤雌发育率明显高于其他两组(P<0.05),囊胚发育率三组之间无显著差异。表明猪卵母细胞体外成熟培养时间延长,体外受精后的囊胚发育率降低,孤雌发育率相应增加。 相似文献
17.
This experiment aimed to study the effect of brilliant cresyl blue (BCB) on in vitro maturation of pig oocytes and the developmental capacity of pig SCNT embryos.The cumulus-oocyte complexes (COCs) were stained with different concentrations of BCB (13,26,39 and 52 μmol/L) for 90 min,and then we divided the COCs into BCB+ and BCB- for in vitro culture 42 to 44 h.The results showed that,with the concentration of BCB increased,the staining rate (20.00%,46.39%,51.66% and 59.03%) raised gradually while the maturation rate of oocytes (74.03%,72.16%,70.53% and 48.61%) reduced,the percentages of oocytes staining by 26 μmol/L BCB for 90 min were higher than that of other groups in staining rate and maturation rate.However,the nuclear maturation rate of BCB+ groups were higher than that of BCB- group.Therefore,26 μmol/L BCB was selected as the most effective concentration dying the oocytes (BCB+),which were used as parthenogenetic activation and nuclear transfer embryos.The cleavage and blastocyst rates of parthenogenetic activation and SCNT embryos in BCB+ group were significantly higher than that of BCB- group (P<0.05),but there were no significant differences between the cleavage and blastocyst rates in the groups of BCB+ and control (P>0.05).Reconstructed embryos derived from the COCs stained with BCB were transferred to five surrogates,and six cloned piglets were obtained from one of the two pregnant pigs.These results showed that COCs stained with BCB was an effective method to select high-quality oocytes,which could improve the efficiency of in vitro embryo production. 相似文献
18.
试验旨在研究亮甲酚蓝(brilliant cresyl blue,BCB)染色对卵母细胞体外成熟及后期胚胎发育潜力的影响。本研究利用13、26、39 、52 μmol/L BCB对成熟培养前的卵丘－卵母细胞复合体(cumulus-oocyte-complexes,COCs)染色90 min,比较各组卵母细胞的着色率、成熟率及孤雌激活胚胎和核移植胚胎的发育情况。结果表明,随着BCB浓度的增加,COCs着色率依次增加(20.00%、46.39%、51.66%和59.03%),但猪卵母细胞体外成熟率逐渐降低(74.03%、72.16%、70.53%和48.61%);不同浓度BCB染色后所得BCB+组卵的成熟率均明显高于BCB-组。试验结果发现,BCB浓度在26 μmol/L时,经染色的COCs既有较高的着色率,且不影响其体外成熟的效率。基于此,研究选取26 μmol/L BCB作为最佳浓度对猪卵母细胞进行染色筛选,然后进行体外培养、孤雌激活及核移植试验。结果显示,筛选的BCB+组卵母细胞的孤雌胚和核移植胚的卵裂率和囊胚率均显著高于BCB-组(P<0.05),而与对照组间无显著差异(P>0.05)。胚胎移植试验挑选BCB+组中发育较好的1-2细胞期重组胚对5头代孕母猪进行了移植,其中2头怀孕,1头顺利产下了6头健康胎儿。综合以上试验结果表明,利用BCB染色可作为一种有效的方法筛选体外成熟质量较高的猪卵母细胞,同时提高胚胎体外生产效率。 相似文献