共查询到17条相似文献，搜索用时 187 毫秒
1.
绵羊卵母细胞的孤雌激活 总被引：2，自引：0，他引：2
本文探讨了不同激活方法对绵羊卵母细胞的孤雌激活和其后的发育。结果表明 ,电激活可以激活绵羊卵母细胞孤雌发育到囊胚 ;Ca2 + 载体A2 3187和CHX组合 ,Ionomycin和 6 DMAP组合可以激活绵羊卵母细胞 ,其卵裂率与电激活相比差异显著。 7%乙醇激活绵羊卵母细胞 7min效果较好。不同场强、不同脉冲次数对绵羊卵母细胞激活都有影响 ,以 1.2kV/cm ,间隔 30 μs和 3次脉冲效果较好。而电激活与化学激活联合可以更好的激活绵羊卵母细胞 相似文献
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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
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猪卵母细胞体外成熟和孤雌激活参数研究 总被引：2，自引：2，他引：0
本研究比较了按不同方法贮藏的培养液对猪卵母细胞体外成熟及成熟后孤雌激活对胚胎发育的影响。此外,还探索了针对初情期前母猪体外成熟卵母细胞和孤雌激活方案。结果如下：①1.4 kv/cm、100 μs、1DC电激活后,卵母细胞死亡率明显高于2.0 kv/cm、30 μs、1DC和2.0 kv/cm、60 μs、1DC处理组,但激活后胚胎卵裂率和囊胚率相似;②使用钙离子载体和6-二甲基氨基嘌呤（Ionomycin+6-DMAP）处理后,胚胎卵裂及囊胚率都明显不如电激活处理;③6次独立试验结果证明：经4℃冷藏和-20℃冷冻保存的培养液用于猪卵母细胞培养,其体外成熟率、孤雌激活后的卵裂率及囊胚发育率无显著差异（P＞0.05）。说明成熟卵在2.0 kv/cm、30 μs、1DC或者 2.0 kv/cm、60 μs、1DC电击参数激活下可以降低死卵率;按本试验设计的电激活方案处理初情期前母猪成熟卵优于化学激活;在-20℃冷冻保存猪卵母细胞成熟液是可行的。 相似文献
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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
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《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
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试验旨在摸索猪卵母细胞孤雌激活的电场强度和脉冲时间,并探索渗透压阶段培养法对孤雌胚胎后期发育的影响。猪卵母细胞成熟培养42~44 h后,分别在电场强度2.1、2.3、2.5 kV/mm和脉冲时间30、60、90 μs的9组电激活参数下进行孤雌激活试验;卵母细胞在2.1 kV/mm和30 μs的参数下进行孤雌激活后,分别培养于渗透压为271、280、290、302 mOsm的PZM-3中,48 h后移入渗透压280 mOsm的PZM-3中继续培养96 h;孤雌胚胎于电激活后先在含2 mmol/L 6-DMAP的PZM-3中培养4~6 h,然后移入不含6-DMAP的PZM-3中继续培养。试验结果表明,电场强度和脉冲时间两个参数间无显著的交互作用(P>0.05),脉冲时间相同条件下,卵裂率在不同电场强度条件下均无显著差异(P>0.05),2.1和2.5 kV/mm的电场强度条件下,脉冲时间为30 μs时的卵裂率显著高于60和90 μs(P<0.05),而2.3 kV/mm电场强度下3个脉冲时间试验组的卵裂率无显著差异(P>0.05),各试验组的囊胚率无显著差异(P>0.05);孤雌胚胎在渗透压为290~310 mOsm的PZM-3中培养48 h,卵裂率得到显著提高(P<0.05),渗透压对囊胚率无显著影响(P>0.05);6-DMAP对孤雌胚胎卵裂率无显著影响(P>0.05),但可以显著提高囊胚率(P<0.05)。结果提示,猪卵母细胞孤雌激活需要较高的电场强度(2.1~2.3 kV/mm)而脉冲时间不宜过长(30 SymbolmA@s);48 h的高渗培养和6-DMAP的辅助激活有助于孤雌胚胎的后期发育。 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.
应用细胞骨架抑制剂细胞松弛素B和秋水仙素处理兔成熟卵母细胞，然后采用电激活的方法激活兔卵母细胞，观察细胞骨架抑制剂对兔卵母细胞孤雌激活和孤雌发育的影响，结果表明：应用不同的电激活液，即甘露醇、Zimmerman氏和山梨醇对兔卵母细胞的孤雌激活效果无显著性差异（P〉0．05）；用7．5μg·mL^-1细胞松弛素B处理卵母细胞后孤雌激活，其2-细胞胚率（82．2％）和囊胚发育率（43．4％）与对照组（76．4％和51．5％）相比无显著性差异（P〉0．05）；用1μg·mL^-1秋水仙素处理兔卵母细胞，兔卵母细胞孤雌激活后的2-细胞胚率（64．9％）和囊胚发育率（23．8％）明显下降，与对照组相比存在显著性差异（P〈0．05）；用细胞松弛素B和秋水仙素共同处理的2-细胞胚率（62．5％）和囊胚发育率（30．9％）与秋水仙素单独处理的结果无显著性差异（P〉0．05）。因此，秋水仙素对兔卵母细胞的孤雌激活影响比CB更大。通过免疫荧光和激光扫描共聚焦显微镜观察发现，细胞骨架抑制剂处理后，经孤雌激活获得的囊胚中，微丝和微管结构未见异常。 相似文献
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采用不同的注射方式结合人工激活方法处理猪卵母细胞,旨在探讨各处理方法对猪ICSI效果的影响。结果表明：假性注射＋未激活处理组卵母细胞的激活率虽然高于无机械刺激＋无激活处理对照组,但明显低于假性注射＋人工激活处理组;精子注射卵母细胞经CaCl2（1.8pL,30mmol/L）、离子霉素（15μmol/L,40min）和电脉冲（场强0.4kV/cm、脉冲时程90μs、1次脉冲）激活处理后,其激活率无显著性差异（P〉0.05）,CaCl2处理组的受精率显著高于离子霉素处理组（P〈0.05）,并与电激活处理组无显著性差异（P〉0.05）;但是,在假性注射组和对照组中,CaCl2处理组的孤雌发育率极显著地低于离子霉素和电激活处理组（P〈0.01）;CaCl2处理组的卵裂率（P〈0.01）和囊胚总细胞数（P〈0.05）显著高于对照组。因此,单纯注射性机械刺激对猪卵母细胞的激活效果较差,有必要进行人工激活;ICSI卵母细胞经CaCl2溶液激活处理后,能够取得较好的ICSI效果,而不显著增加其孤雌发育率。 相似文献
12.
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
13.
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
14.
SR Lee J-W Kim BS Kim D-H Yoo YS Park T-H Lee J-H Ha B-H Hyun ZY Ryoo 《Reproduction in domestic animals》2009,44(5):740-744
In this study, we investigated parthenogenetic induction of canine oocytes by electrical stimulation following Ca-EDTA treatment. Oocyte maturation, parthenogenetic development, and cleavage rate in canine after various electrical stimulations (1.5, 1.8, 2.1 kV/cm) for 50 μs with single DC pulse following 1 mM Ca-EDTA treatment were investigated. In oocyte activated electrically at the voltage of 1.5 kV/cm after 1 mM Ca-EDTA treatment, the rate of pronucleus and two-cell was 4.1% and 2.7%, respectively. Although electrical stimulation could parthenogenetically induce immature oocyte to cleavage stage, degeneration rate in all experimental groups was more than 60%. This means that electrical stimulation after Ca-EDTA treatment could cause canine oocytes to be degenerated. However, two-cell in canine oocyte by parthenogenesis was for the first time induced. Therefore, we suggested that electrical stimulation for canine oocytes could induce parthenogenetically early embryonic cleavage. This result can be used as a basic data for parthenogenesis study in canine. Also, to perform more developed embryonic development, further study to parthenogenesis in canine need to be developed. 相似文献
15.
A George RA Shah R Sharma P Palta SK Singla RS Manik MS Chauhan 《Reproduction in domestic animals》2011,46(3):444-447
Parthenogenetic activation using zona‐free oocytes offers an alternative model that could be applied to develop protocols for the activation of reconstructed embryos for cloning. The aim of this study was to compare the efficacy of different methods for the activation of zona‐free buffalo oocytes in terms of their effects on the developmental competence of parthenogenetic embryos. The effects of zona removal on parthenogenetic activation and in vitro developmental competence of metaphase II oocytes were also examined. All activation methods were followed by incubation of 2 mm 6‐dimethylaminopurine (6‐DMAP) for 4 h. Out of three different pulse strengths (1.2, 2.1 or 3.3 kV/cm) used, 2.1 kV/cm resulted in the highest blastocyst rate (25.3%). On comparing different chemical agents and electric pulse, highest blastocyst rate was observed for calcium ionophore (CaI) (28.6%) followed by ethanol (25.0%), electric pulse (22.5%) and combined CaI and ethanol treatment (16.7%) although differences among them were not significant. Furthermore, a significantly reduced developmental potential was observed in zona‐free oocytes when compared to zona‐intact ones up to the blastocyst stage (44.3% vs 27.1%). In conclusion, zona‐free buffalo oocytes can be successfully activated for parthenogenetic development using chemical or electrical stimulation. Out of different agents examined, CaI followed by 6‐DMAP resulted in the highest blastocyst rate. 相似文献
16.
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
17.
D Kwon IM Saadeldin SJ Kim SJ Park JT Kang HJ Park JH Moon OJ Koo G Jang BC Lee 《Reproduction in domestic animals》2014,49(6):995-999
Modifying electrical activation conditions have been used to improve in vitro embryo production and development in pigs. However, there is insufficient information about correlations of porcine embryo development with oocyte pre‐ and post‐activation conditions. The purpose of this study was to compare the developmental rates of porcine oocytes subjected to different mannitol exposure times, either pre‐ or post‐electrical activation, and to elucidate the reason for the optimal mannitol exposure time. Mannitol exposure times around activation were adjusted as 0, 1, 2 or 3 min. Blastocyst development were checked on day 7. Exposure of oocytes to mannitol for 1 or 2 min before electrical activation produced significantly higher blastocyst rates than exposure for 0 or 3 min. There was no significant difference in blastocyst rates when activated oocytes were exposed to mannitol for 0, 1, 2 or 3 min after electrical activation. While exposure of oocytes to mannitol for 1 min pre‐ and 3 min post‐activation showed significantly higher blastocyst development than 0 min pre‐ and 0 min post‐activation. It also showed higher maintenance of normal oocyte morphology than exposure for 0 min pre‐ and 0 min post‐activation. In conclusion, exposure of oocytes to mannitol for 1 min pre‐ and 3 min post‐activation seems to be optimal for producing higher in vitro blastocyst development of porcine parthenogenetic embryos. The higher blastocyst development is correlated with higher maintenance of normal morphology in oocytes exposed to mannitol for 1 min pre‐ and 3 min post‐activation. 相似文献