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兔核移植胚胎的克隆研究 总被引:12,自引:0,他引:12
本研究改进了兔受体卵母细胞的去核程序及供体卵裂球的处理方法,并对核移植胚的体外克隆、继代克隆及克隆胚的体内发育进行了试验。结果表明:卵龄对受体卵母细胞的去核具有显著的影响,卵龄为15~18h的去核率为100%(45/45),显著高于13~14h的46.6%(7/15),P<0.01。DNA合成抑制剂Aphidicolin处理16-细胞期卵裂球后,重组胚的囊胚发育率54%(20/37)高于未处理组的45%(14/31),P<0.05。从2枚16-细胞供体胚分别获得来自一个供体胚的9枚和7枚核移植克隆囊胚,囊胚发育率为51.6(16/31)。然而第一次继代核移植胚的囊胚发育率仅为11.5%(6/52)。16-及32-细胞期卵裂球的重组胚可发育至产仔,共获2窝8只仔兔。其中1只、2只、2只和3只仔兔分别来自4个供体胚。 相似文献
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影响核移植克隆胚胎效率的因素分析刘林,安民(北京农业大学动物科技学院100094)一、前言*采用核移植技术扩增遗传上优秀的哺乳动物胚*胎,对于提高动物生产力具有巨大潜力。近年来哺乳动*物核移植技术取得了很大进展。已经获得了包括大家*畜在内的多种动物的... 相似文献
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供体细胞对猪体细胞克隆胚胎早期发育的影响 总被引:1,自引:1,他引:1
以中国农业大学实验用小型猪香猪胎儿成纤维细胞、成年耳成纤维细胞和颗粒细胞3种细胞系为供体细胞进行核移植。比较了血清饥饿法和接触抑制法处理胎儿成纤维细胞诱导进入G0/G1期的效率,发现二者差异不显著(P〉0.05),血清饥饿2d和4d差异不明显,同样接触抑制2d和4d差异也不显著(P〉0.05)。系统研究了影响克隆胚胎发育的供体因素:血清饥饿与否、细胞形态、细胞类型及个体差异等,结果表明:血清饥饿处理对克隆胚的早期发育没有明显的促进作用;圆形光滑细胞有利于细胞融合,对早期发育无显著影响(P〉0.05);不同个体、不同类型的供体细胞对克隆胚囊胚发育率有一定的影响。 相似文献
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对影响小鼠胚胎干细胞(Embryonic stem cells,ES细胞)培养、克隆、分离、传代效果的因素进行了探索研究。应用223枚昆明白小鼠胚胎和20枚129品系小鼠胚胎的研究结果表明,129品系小鼠胚胎比昆明白小鼠胚胎更适合作为ES细胞建系的材料,两者FS出现率差异显著(P<0.05);以DMEM+10%NBS+10%FCS为基础培养液,分别加入LIF、胰岛素、LIF+SCF,极显著提高昆明白小鼠胚胎贴壁率,ICM生长率及F1、F2出现率(P<0.01),而在DMEM+10%NBS+10%FCS+LIF+SCF为培养液,得到昆明白小鼠胚胎最高贴壁率、ICM生长率及传代率;4dpc胚胎传代情况显著好于3.5dpc胚胎(P<0.05)。 相似文献
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<正>动物克隆胚胎安全生产是指在动物克隆胚胎生产过程中,为了避免人员人身伤害、财产损失以及胚胎无效生产等生产事故的发生,采取一系列事故预防和控制措施,保障人员安全与健康、设施设备免受损失、克隆胚胎高效生产的相关活动。克隆胚胎安全生产主要涉及前期准备、细胞培养、卵母细胞收集、卵母细胞体外成熟、核移植、胚胎体外培养六大环节,每个环节均是保障克隆胚胎顺利生产必不可少的条件,而每个环节都会涉及一些具体的安全问题。 相似文献
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对昆明小鼠和BALB/c小鼠2种胚胎的研究表明,2种品系小鼠胚胎均可用作胚胎干细胞(ES)分离建系的材料,两者在ES分离、传代上无显著差异(P〉0.05);大鼠心肌细胞条件培养液与添加LIF的ES常规培养液相比,显著提高了小鼠ES细胞F1、F2出现率(P〈0.05);采用低浓度消化液使形成ES克隆的比率分别从14%、16%提高到35%、32%,使ES传到第5代的比率分别从1.7%、0提高到5%、7.1%;而采用连续消化法使形成ES克隆的比率从15%、17%提高到40%、50%,使ES传到第5代的比率从0、1%提高到10%、20%;对ES进行伊红染色、核型分析、AKP染色及体外分化能力检测,证实所分离的ES符合小鼠ES的一系列特征。 相似文献
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用8~12周龄BALB/c公鼠采取皮肤移植和脾细胞腹腔注射2种方式免疫同日龄纯系BALB/c母鼠,制备H-Y抗血清。经精子微量细胞毒性试验,筛选的抗血清用于胚胎体外培养,以鉴定附植前小鼠胚胎上H-Y抗原的表达。选用8cell-早期囊胚进行体外培养,分别于5~6、18和24h观察胚胎发育情况。5~6h后,H-Y抗血清培养的胚胎退化率高达40%,且发育迟缓,与自然性比例差异不明显(P〉0.05)。正常 相似文献
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Xiao-ying He Li-bing Ma Xiao-ning He Wan-tong Si Yue-Mao Zheng 《Journal of veterinary science (Suw?n-si, Korea)》2016,17(2):145-152
Previous studies have established a bovine mammary gland epithelia cells in vitro model by the adenovirus-mediated telomerase (hTERT-bMGEs). The present study was conducted to confirm whether hTERT-bMGEs were effective target cells to improve the efficiency of transgenic expression and somatic cell nuclear transfer (SCNT). To accomplish this, a mammary-specific vector encoding human lysozyme and green fluorescent protein was used to verify the transgenic efficiency of hTERT-bMGEs, and untreated bovine mammary gland epithelial cells (bMGEs) were used as a control group. The results showed that the hTERT-bMGEs group had much higher transgenic efficiency and protein expression than the bMGEs group. Furthermore, the nontransgenic and transgenic hTERT-bMGEs were used as donor cells to evaluate the efficiency of SCNT. There were no significant differences in rates of cleavage or blastocysts or hatched blastocysts of cloned embryos from nontransgenic hTERT-bMGEs at passage 18 and 28 groups (82.8% vs. 81.9%, 28.6% vs. 24.8%, 58.6% vs. 55.3%, respectively) and the transgenic group (80.8%, 26.5% and 53.4%); however, they were significantly higher than the bMGEs group (71.2%, 12.8% and 14.8%), (p < 0.05). We confirmed that hTERT-bMGEs could serve as effective target cells for improving development of somatic cell cloned cattle embryos. 相似文献
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从不同发情方式、不同胚胎类型、不同胚龄以及不同季节来探讨影响水牛体细胞克隆胚胎移植的效率。结果显示,自然发情与同期发情方法对水牛体细胞克隆胚胎的移植受胎率有显著影响。鲜胚和冻胚移植后受胎率没有显著差异,其受胎率分别是9.79%、14.5%。在移植中,不论是鲜胚还是冻胚,6日龄胚胎移植受胎率最高。而且不同日龄(5、6、7、8~10d)克隆胚胎移植受胎率差异不显著(鲜胚P=0.260 7,冻胚P=0。065 2)。克隆胚胎在秋季移植时受胎率最高,受胎率是16.06%,比其他季节高,且差异显著(P<00.05).其次是春季,但夏季和冬季克隆胚胎移植后受胎率差异不显著(P>0.05)。结果表明,水牛体细胞克隆胚胎在胚胎移植中是可行的,但效率比较低。 相似文献
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Preimplantation embryos from mice and cattle were exposed to bluetongue virus in vitro to determine whether the virus would replicate in these early embryos and, if so, what pathologic consequences would ensue. A high proportion of zona pellucida-free, 2-cell embryos and morulae from mice, and morulae from cattle became infected. The infection was rapidly cytopathic in embryos from both species. Indirect immunofluorescence was used to demonstrate accumulation of virus antigen in the blastomeres of these embryos. The zona pellucida of both murine and bovine embryos provided effective protection from virus present in culture fluid. 相似文献
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由于在核移植试验中,所用的水和化学物质都不可避免的会被一些金属离子轻微污染,造成胚胎内抗氧化物和过氧化物之间难以保持平衡,从而导致胚胎发育率降低,本试验以此为出发点,探讨了在延边黄牛体细胞核移植重组胚早期培养液中添加乙二胺四乙酸钠(EDTA-Na)和柠檬酸钠(sodium citrate)2种金属螯合剂类抗氧化剂,对其后续发育的影响,以期筛选出最佳的体外培养条件。结果表明:适合延边黄牛体细胞核移植重组胚后期发育的EDTA-Na和柠檬酸钠的最佳浓度分别为50μmol/L和0.6mmol/L。 相似文献
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Ting-Yu ZHANG Jian-Jun DAI Cai-Feng WU Xiao-Long GU Liang LIU Zhi-Qiang WU Yi- Ni XIE Bin WU Hui-Lan CHEN Yao LI Xue-Jin CHEN De-Fu ZHANG 《Animal Science Journal》2012,83(2):103-110
To optimize somatic cell nuclear transfer (SCNT) procedures in mini-pigs, the present study was designed to examine the effects of donor cell types and aphidicolin (APC) treatment on in vitro development of reconstructed embryos. Oviduct epithelial cells (OEC), ear fibroblast cells (EFC) and cumulus cells (CC) derived from mini-pigs were treated with serum starvation only or serum starvation followed by treatment of 0.1 µg/mL APC. The reconstructed embryos were cultured for 7 days to evaluate their developmental competency. Cleavage and blastocyst formation rates of reconstructed embryos derived from the OEC by APC treatment were significantly higher than the serum starvation (61.82% vs. 56.25%, 24.55% vs. 17.86%; P < 0.05). The cleavage rate from the EFC was significantly increased by APC treatment compared to serum starvation only (63.36% vs. 57.01%; P < 0.05). In the ooctyes with the CC, the reconstructed embryos could yield high blastocyst formation rate by APC treatment (29.63%; P < 0.05). In the presence of APC, CC gave rise to the highest cleavage and blastocyst formation rates among the three cell types. Therefore, our results suggest that treatment of CC with serum starvation plus APC prior to nuclear transfer is more suitable in SCNT of mini-pigs. 相似文献
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Parthenogenetic activation is an important factor in successful production of cloned mammals. Because it has been reported that aged oocytes are more sensitive to parthenogenetic activation than young oocytes, the present study examined the effects of oocyte aging on the in vitro and in vivo developmental potential of nuclear-transferred (NT) mouse oocytes receiving cumulus cells. The potentials of young NT oocytes (14 h after human chorionic gonadotrophin [hCG] injection) to develop into blastocysts was, however, significantly higher than that of aged oocytes (20 h after hCG injection; 16% vs 6%). When the nuclei of NT oocytes at the 2-cell stage were fused with enucleated fertilized 2-cell embryos, the potentials of the serial NT embryos to develop into blastocysts were no different for both young and aged oocytes (74% vs 74%). Live young, however, were obtained only after transfer of serial NT blastocysts developed from young NT oocytes (2%). In contrast to a report using embryonic nuclei as the nuclear donors, the results of the present study indicate that young oocytes are superior to aged oocytes as a source of recipient cytoplasm for mouse somatic cell cloning. 相似文献
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聚乙烯醇、聚乙烯吡咯烷酮和牛血清白蛋白对牛卵母细胞成熟及体细胞克隆胚胎发育的影响 总被引:1,自引:0,他引:1
为了比较不同浓度的聚乙烯醇(PVA)、聚乙烯吡咯烷酮(PVP)、牛血清白蛋白(BSA)对牛卵母细胞体外成熟率、重组胚卵裂率和发育率的影响,试验采用抽吸法回收卵母细胞,用不同的无血清成熟液进行卵母细胞的体外成熟培养,然后对成熟卵母细胞进行核移植、融合、激活以及胚胎的体外培养.结果表明,适合延边黄牛卵母细胞成熟及体细胞克隆胚胎发育的PVA、PVP和BSA的最佳浓度分别是1.0 mg/mL,0.3%和0.4%. 相似文献