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Effects of chemical activation and season on birth efficiency of cloned pigs 被引量:2
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作者 MA YuFang1,2,LI Yan3,WEI HengXi1,LI QiuYan3,FANG Rui1,ZHAO Rui3,ZHANG Kun1,XUE Kai1,LOU YanKun1,DAI YunPing3,LIAN LinSheng2 & LI Ning1 1 State Key Laboratory of Agrobiotechnology,College of Biological Sciences,China Agricultural University,Beijing 100193,China 2 College of Animal Science and Technology,China Agricultural University,Beijing 100193,China 3 Beijing Genprotein Biotechnology Company,Beijing 100193,China 《Science China(Life Sciences)》 SCIE CAS 2009年第7期657-664,共8页
The effects of chemical activation on birth efficiency of cloned pigs were studied by investigating the developmental process from porcine oocyte activation to birth of cloned pigs.Three different activation methods w... The effects of chemical activation on birth efficiency of cloned pigs were studied by investigating the developmental process from porcine oocyte activation to birth of cloned pigs.Three different activation methods were used:(i) Electroporation(Ele);(ii) Ele followed by incubation with 6-dimethylaminopurine(6-DMAP);and(iii) Ele followed by a treatment with cycloheximide(CHX).In experiment 1,the rates of cleavage,developmental rates and cell number of porcine parthenogenetic(PA) embryos were investigated in the three treatment groups.In experiment 2,NT embryos produced by the three different activation treatments were compared for the rates of cleavage,development and cell number.Finally,the effects of Ele and Ele+CHX activation methods on birth efficiency of cloned pigs were compared.The activated oocytes treated by combination activation generally showed a higher(P<0.05) blastocyst rate and produced more expanded blastocysts than oocytes activated with Ele.The rates of cleavage and total cell number of parthenotes were not significantly different.Parthenogenetic embryos activated with 6-DMAP developed into blastocyst and expanded blastocyst stages at a significantly(P<0.05) higher rate than those treated with Ele,but the developmental capability was dramatically decreased in NT embryos.With the CHX activation method,the NT embryo blastocyst rate was substantially(P<0.05) increased although the production of expanded blastocysts was not significantly different from that by the other two methods.The birth rate of cloned pigs increased in the CHX group,though the rate was not significantly different from Ele.The effects of season on developmental rate of the porcine PA embryos and birth rate of cloned pigs were also examined in our study.Porcine oocytes collected in the spring had higher developmental capabilities than those collected in the winter.However,no difference in birth rate of the cloned pigs was found between the oocytes collected in the two seasons.The results obtained from PA and NT embryos,following different activation methods,were inconsistent,suggesting that activation mechanisms are dissimilar in PA and NT embryos.Although the chemical activation in our study leads to an elevation of the blastocyst rate,it does not improve the oocyte's molecular programming and so does not significantly improve the efficiency of producing cloned pig births. 展开更多
关键词 parthenogenetic activation chemical activation nuclear transfer pig season
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猪体外受精胚胎、孤雌激活胚胎以及体细胞核移植胚胎的体外培养 被引量:10
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作者 张运海 潘登科 +4 位作者 孙国杰 孙秀柱 李燕 戴蕴平 李宁 《中国农业科学》 CAS CSCD 北大核心 2007年第3期588-593,共6页
目的本研究系统比较了体外受精胚(IVFEs)、孤雌激活胚(PAEs)以及体细胞核移植胚(NTEs)的发育率和囊胚质量,同时还比较了PZM-3和NCSU-23两种培养基培养对PAEs以及NTEs发育的影响。方法利用体细胞核移植技术、体外受精技术和孤雌激活技术... 目的本研究系统比较了体外受精胚(IVFEs)、孤雌激活胚(PAEs)以及体细胞核移植胚(NTEs)的发育率和囊胚质量,同时还比较了PZM-3和NCSU-23两种培养基培养对PAEs以及NTEs发育的影响。方法利用体细胞核移植技术、体外受精技术和孤雌激活技术分别生产猪的NTEs、IVFEs和PAEs,开展体外培养。结果(1)IVFEs、PAEs和NTEs的卵裂率分别为72.0%,66.0%和63.5%,各组间没有显著差异(P>0.05);囊胚形成率分别为11.0%,22.6%和16.9%,也不存在明显不同(P>0.05);然而,在囊胚质量,即ICM细胞数/总细胞数的比例上,IVFEs和NTEs均优于PAEs(0.448%,0.356%vs 0.122%,P<0.05)。(2)对PAEs来讲,以PZM-3为基础培养基时囊胚率明显高于以NCSU-23为基础培养基时的处理组(35.4%vs.22.6%,P<0.05);而对NTEs而言,两种培养基在支持胚胎形成囊胚方面的效果相当(26.6%vs.21.1%,P>0.05),虽然PZM-3培养时胚胎卵裂率显著低于NCSU-23培养组(47.5%vs.63.5%,P<0.05)。结论(1)PAEs的囊胚质量在3种来源的胚胎中最差;(2)对PAEs理想的培养基,当用于NTEs培养时却不一定是最佳的选择。 展开更多
关键词 体外受精 体细胞核移植 孤雌激活 体外培养
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不同类型绵羊胚胎体外发育率的比较研究
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作者 潘晓燕 杨梅 +4 位作者 王正朝 汪立琴 陈童 郭志勤 王锋 《南京农业大学学报》 CAS CSCD 北大核心 2009年第2期112-117,共6页
分别利用孤雌激活技术、体外受精技术及体细胞核移植技术生产不同类型的体外胚胎,比较不同类型胚胎的体外发育率和囊胚细胞总数间的差异,研究季节、卵巢保存条件和培养液对孤雌激活胚(PAE)和体细胞核移植胚(SCNTE)发育能力的影响... 分别利用孤雌激活技术、体外受精技术及体细胞核移植技术生产不同类型的体外胚胎,比较不同类型胚胎的体外发育率和囊胚细胞总数间的差异,研究季节、卵巢保存条件和培养液对孤雌激活胚(PAE)和体细胞核移植胚(SCNTE)发育能力的影响。结果表明:PAE、SCNTE和体外受精胚(IVFE)的卵裂率没有显著差异;PAE的囊胚率(23.4%)显著高于SCNTE(18.2%);SCNTE的囊胚细胞数(120)显著多于PAE的囊胚细胞数(96),而与IVFE的囊胚细胞数(107)没有显著差异。繁殖季节卵巢PAE和SCNTE的发育率显著高于非繁殖季节。卵巢在14~18℃保存15~17h,没有影响PAE的体外发育,却显著降低了SCNTE的卵裂率,且没有得到囊胚。SOFaa和CR1aa培养基对PAE和SCNTE的早期发育没有显著影响。由此可见,季节和卵巢的保存方式对绵羊胚胎的体外发育率有显著影响,且不同类型胚胎的质量也存在显著差异。 展开更多
关键词 绵羊 孤雌激活 体细胞核移植 体外受精 季节 卵巢 培养液
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亮甲酚蓝对猪卵母细胞体外成熟及核移植胚胎发育能力的影响 被引量:1
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作者 徐国旗 华再东 +1 位作者 许生成 郑新民 《中国畜牧兽医》 CAS 北大核心 2016年第3期754-760,共7页
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、52μmol/L BCB对成熟培养前的卵丘-卵母细胞复合体(cumulus-oocyte-complexes,COCs)染色90min,比较各组卵母... 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、52μmol/L BCB对成熟培养前的卵丘-卵母细胞复合体(cumulus-oocyte-complexes,COCs)染色90min,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26μmol/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26μmol/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。 展开更多
关键词 卵母细胞 体外成熟 亮甲酚蓝染色 孤雌激活 核移植
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