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1.
采集中国大白兔的桑椹胚共545枚,采用OPS法,配制VSⅠ和VSⅡ两种冷冻液,分别冷冻兔的桑椹胚265枚和280枚,解冻后胚胎回收率分别为93.6%(248/265)和92.8%(260/280),两者差异不显著(P>0.05);解冻后胚胎形态良好率分别为95.2%(236/248)和94.2%(245/260),两者差异显著(P<0.05)。将解冻后形态良好的210枚桑椹胚分别移植到15只受体母兔体内,结果有8只母兔妊娠,妊娠率为53.3%(8/15),妊娠受体内移植胚胎数为110枚,产仔数占移植胚胎数的33.6%(37/110)。用VSⅠ冷冻的100枚桑椹胚移植到7只受体母兔体内,妊娠率为57.1%(4/7),产仔率为35.7%(20/56);用VSⅡ冷冻的110枚桑椹胚移植到8只受体母兔体内,妊娠率为50.0%(4/8),产仔率为31.5%(17/54)。无论妊娠率还是产仔率,VSⅠ都高于VSⅡ,但差异均不显著(P>0.05)。  相似文献   

2.
采用2种不同浓度的冷冻液(VSⅠ和VSⅡ),通过OPS法玻璃化冷冻兔的胚胎。结果表明:冷冻兔的桑椹胚641枚,解冻后胚胎总回收率为93.8%(601/641),形态结构良好率为94.5%(568/601),透明带完好率为100%(601/601)。VSⅠ和VSⅡ冷冻的胚胎解冻后回收率分别为93.8%(305/325)和93.7%(296/316),两者差异不显著(P>0.05);形态良好率分别为95.1%(290/305)和94.0%(278/296),前者显著地高于后者(P<0.05)。将解冻后形态结构良好的桑椹胚415枚移植到29只受体母兔体内,总妊娠率为51.7%(15/29),产仔率为34.5%(61/177)。VSⅠ妊娠率为53.3%(8/15),产仔率为36.8%(35/95);VSⅡ妊娠率为50.0%(7/14),产仔率为31.7%(26/82)。无论妊娠率还是产仔率,VSⅠ都高于VSⅡ,但差异不显著(P>0.05)。  相似文献   

3.
为了研究牛早期胚胎取样对移植妊娠率的影响,本试验从胚胎类型、胚胎质量、胚胎发育阶段、取样细胞数及体外放置时间五个方面进行分析。结果表明,整胚与取样性鉴胚胎之间移植妊娠率差异不显著(P0.05);取样后1级胚胎的移植妊娠率极显著高于取样后的2级胚胎(P0.01);扩张囊胚的移植妊娠率显著高于早期囊胚和桑葚胚的移植妊娠率(P0.05)。1级和2级胚胎均在取样4~6个细胞时,获得最高移植妊娠率,但与其他组别差异均不显著(P0.05)。随着胚胎在体外放置时间的延长,移植妊娠率呈现下降的趋势。  相似文献   

4.
绵羊玻璃化冷冻胚胎直接移植试验研究   总被引:1,自引:0,他引:1  
应用EFS40玻璃化液对6.5~7日龄的绵羊胚胎进行玻璃化冷冻及解冻后直接移植试验.结果:桑椹胚、囊胚冷冻解冻后移植的妊娠率分别为37.50%(3/8)和54.55%(6/11),胚胎存活率分别为33.33%(3/9)和50.00%(6/12),差异均不显著(P>0.05);胚胎解冻后用0.5 mol/L蔗糖脱防冻剂与直接用胚胎存放液脱除防冻剂的妊娠率分别为44.44%(4/9)和50.00%(5/10),胚胎存活率分别为40.00%(4/10)、45.45%(5/11)差异不显著(P>0.05);10枚解冻后的胚胎细管内脱防冻剂后,直接装管移植给8只受体,妊娠率为50.00%(4/8),胚胎成活率为40.00%(4/10),与同期常规冷冻解冻组相比无显著差异(P>0.05).  相似文献   

5.
统计分析了河北省2007~2009年度胚胎移植妊娠情况。结果表明:1)胚胎级别对妊娠率有显著影响(43.02%∶36.65%,P0.01);2)胚胎不同发育阶段对妊娠率没有显著影响,但早期囊胚妊娠率为44.34%,高于其他组(P0.05);3)胚胎冷冻(59.38%∶50.00%)与胚胎取样(54.69%∶51.35%)对胚胎的妊娠率均有一定的影响,但差异均不显著(P0.05);4)春季和冬季的妊娠率为36.42%和37.74%,显著低于夏季移植妊娠率45.82%(P0.05)与秋季无显著差异(P0.05)。夏季和秋季胚胎移植妊娠率差异不显著(45.82%∶42.60%,P0.05);5)受体牛营养状况对移植妊娠率影响显著(P0.01)。  相似文献   

6.
为探索出一条能够提高牛胚胎移植妊娠率的有效途径,本研究分别将胚胎滋养层和胚胎块(均为雄性)分别与性别鉴定冻胚共移植,得出妊娠率分别为51.2%和62.5%,均高于对照组(不加雄性胚胎块)43.8%的妊娠率(P>0.05);两个试验组间无显著差异。鲜滋养层和鲜胚胎块分别与性别鉴定鲜胚共移植妊娠率高于对照组21.4个百分点(69.2%vs 47.8%)(P>0.05)和12.2个百分点(60.0%vs 47.8%)(P>0.05)。  相似文献   

7.
对影响后的性别鉴定胚胎移植妊娠率的因素进行了分析。结果表明,性别鉴定胚胎的鲜胚和冻胚移植妊娠率均低于非性别鉴定胚胎,但二者差异不显著(P>0.05),这表明性别鉴定胚胎在生产中应用是可行的。随着胚胎细胞取样数的增多,其移植妊娠率逐渐下降,且冷冻带来的损伤明显增大;性别鉴定胚胎在体外放置时间小于5 h的妊娠率比大于5 h的高,且差异极显著(P<0.01);性别鉴定胚胎移植时受体的最佳移植时间在发情后的7.5~8.5 d;繁殖力好的受体牛移植妊娠率比繁殖力差的受体牛移植妊娠率高,且差异极显著(P<0.01)。  相似文献   

8.
探讨了胚胎分割前后冷冻和不同时期的冷冻胚胎对水牛胚胎分割效果的影响。结果:冷冻后分割组的分割成功率与分割后冷冻组差异不显著(71.39%vs 71.43%,P>0.05),但囊胚冷冻后分割组的半胚发育率显著高于分割后冷冻组(58.33%vs 47.14%,P<0.05);扩张囊胚的冷冻存活率显著高于孵化囊胚(94.84%vs 79.01,P<0.05),但囊胚与孵化囊胚差异不显著(92.67%vs 79.01%,P>0.05),囊胚、扩张囊胚和孵化囊胚冷冻胚胎的分割成功率(70.76%,73.22%,70.92%)及半胚发育率差异均不显著(53.07%,51.49%,57.99%)(P>0.05)。上述结果说明,水牛囊胚冷冻后分割比分割后冷冻能获得更高的半胚发育率;水牛未扩张囊胚和扩张囊胚的冷冻存活率比孵化囊胚高,且这3个时期的冷冻胚胎均适合分割。  相似文献   

9.
不同因素对牛体外受精效果的影响   总被引:1,自引:0,他引:1  
比较公牛个体、精子密度、精卵比例、受精前机械脱除卵丘细胞及胚胎培养液体积对体外受精效果的影响。结果表明:不同公牛个体的精子受精的卵裂率差异不显著(P>0.05),但囊胚率受到极显著的影响(52.24%vs28.13%,P<0.01);精子密度在(0.5~1.5)×106/mL间的卵裂率和囊胚率差异都不显著(P>0.05),但精子密度在0.25×106个/mL时的卵裂率显著低于其他3个试验组(52.17%vs91.23%,P<0.05);受精时,精卵比控制在2 000~10 000∶1之间,受精卵的卵裂率、囊胚率均无差异(P>0.05);受精前机械脱除卵丘细胞对受精效果没有影响(45.21%vs36.46%,P>0.05);每枚胚胎培养液体积在2.5~10μL之间的群体培养,对受精胚的发育没有显著影响(51.39%vs35.06%,P>0.05)。  相似文献   

10.
试验选用72头杜长大断奶仔猪,随机分为3组,每组3个重复,每个重复8头。试验分为对照组Ⅰ(基础日粮含100 mg/kg无机铁)、对照组Ⅱ(基础日粮+50 mg/kg无机铁)、试验组(基础日粮+50 mg/kg甘氨酸铁),试验结果表明:对照组Ⅰ、对照组Ⅱ和试验组3个组之间差异不显著,试验组的日增重比对照组Ⅰ和对照组Ⅱ分别提高2.67%(P>0.05)和2.18%(P>0.05);从料肉比来看,试验组和对照组Ⅰ差异显著,但与对照组Ⅱ相比差异不显著。分别比对照组Ⅰ和对照组Ⅱ降低1.4%(P<0.05)和0.7%(P>0.05);在腹泻率方面,试验组与对照组Ⅰ之间差异显著(P<0.05),但与对照组Ⅱ之间差异不显著(P>0.05)。试验组比对照组Ⅰ和对照组Ⅱ分别降低93.3%和83.3%;育成率3组之间差异不显著;血清微量元素铁增加的趋势试验组均高于对照组Ⅰ和对照组Ⅱ,组间差异均不显著(P<0.05);试验组粪中微量元素铁的含量比对照组Ⅰ和Ⅱ分别降低30.02%和30.47%,且试验组和对照组Ⅰ、对照组Ⅱ之间差异极显著(P<0.01),但对照组Ⅰ和对照组Ⅱ之间差异不显著(P>0.05)。由此可见,在基础日粮中添加50 mg/kg(含二价铁)甘氨酸铁不仅能提高断奶仔猪生产性能,还能减少粪中微量元素铁的排放量。  相似文献   

11.
The effects of different cell monolayers on in vitro development of early bovine embryos derived from in vitro maturation and fertilization were examined in this study. Early embryos (four to eight cells) were randomly allocated to bovine granulosa cell (GC), oviductal cell (OC), or uterine cell (UC) monolayers in Exp. 1 and to GC, skin cell (SC; from 10-d-old chicken embryos), testicular cell (TC; from 10-d-old mouse), and liver cell (LC; from 10-d-old chicken embryos) monolayers in Exp. 2, and cultured for 6 d at 38.6 degrees C in a humidified atmosphere of 5% CO2 in air. The culture medium was 12.5 mM HEPES TCM 199 supplemented with 1% calf serum and 1 mM sodium pyruvate. In Exp. 1, the percentage of four- to eight-cell embryos that developed to blastocysts on GC, OC, and UC monolayers was 26.9 (28/104), 37.5 (39/104), and 39.2 (40/102), respectively. In Exp. 2, the percentage of four- to eight-cell embryos that developed to blastocysts on GC, SC, TC, and LC monolayers was 53.3 (40/75), 42.9 (33/77), 49.3 (37/75), and 44.3 (35/79), respectively. There were no significant differences in development among groups in either experiment.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

12.
牛体外受精早期胚胎与小鼠胎仔成纤维细胞共培养的研究   总被引:1,自引:0,他引:1  
探讨了人工合成培养液CR1aa和小鼠胎仔成纤维细胞对牛体外受精早期胚胎体外发育的影响。结果表明,牛体外受精卵在CR1aa液中的卵裂率达76.2%,8细胞胚的比率达44.8%。小鼠胎仔成纤维细胞能够显著促进牛体外受精的早期囊胚以上胚胎的发育。牛体外受精后第5、6天的早期胚胎分别与小鼠胎仔成纤维细胞共培养,在受精后第7天发育至囊胚以上的比率分别达19.8%和24.6%;受精后第8天,孵化的囊胚比例分别达5.2%和7.5%。实验表明,受精后第5、6天的牛体外受精早期胚胎与小鼠胎仔成纤维细胞共培养,可显著提高扩张囊胚和孵化囊胚数量。小鼠成纤维细胞对胚胎发育的支持作用取决于胚胎发育阶段  相似文献   

13.
模拟发情期(0~6 d)母牛外周血雌激素和孕酮浓度的变化,添加17β-雌二醇和孕酮至TCM-199中,体外培养间情期牛输卵管上皮细胞(BOECs),同时观察了BOECs分泌蛋白(BOEP)、发情期母牛输卵管冲洗蛋白(BOP)和小牛输卵管冲洗蛋白(COP)对小鼠胚胎发育的影响。结果表明:①类固醇激素可以调节和启动体外培养的间情期牛输卵管上皮细胞的分泌活动;②BOEP比BOP和COP能极显著地促进胚胎的分裂和发育(P<0.001);③与对照组(未添加外源蛋白质,只添加胎牛血清)相比,BOEP的囊胚形成率极显著低于对照组(P<0.01),可能是缺乏某些分子量低于10 kD蛋白因子的协同作用,同时30~56kD分泌蛋白的存在,也可能参与了抑制和阻碍胚胎分裂和发育的过程。  相似文献   

14.
This study was conducted to investigate the developmental capacity of domestic cat-bovine reconstructed embryos via interspecies somatic cell nuclear transfer (iSCNT) and to observe the mitochondrial DNA (mtDNA) content of the iSCNT embryos. The iSCNT embryos were generated using mixed-breed domestic cat fibroblasts as donor cells and enucleated bovine oocytes as the recipient cytoplasm. When the developmental capacities of iSCNT embryos and parthenogenic bovine embryos were compared, there was no difference (P>0.05) in the rates of cleavage and development to the 8-cell stage (86.6 vs. 84.0% and 32.2 vs. 36.2%, respectively). However, in contrast to development of parthenogenic embryos to the morula and blastocyst stages, no iSCNT embryos (0/202) developed beyond the 8-cell stage. For mtDNA analysis, iSCNT embryos at the 1-cell, 2-cell, 4-cell and 8-cell stages were randomly selected. Both cat and bovine mtDNA quantification analysis were performed using quantitative PCR. The levels of both cat and bovine mtDNA in cat-bovine iSCNT embryos varied at each stage of development. The cat mtDNA concentration in the iSCNT embryos was stable from the 1-cell to 8-cell stages. The bovine mtDNA in the iSCNT embryos at the 8-cell stage was significantly lower than that at the 4-cell stage (P<0.05). No difference in the proportions of cat mtDNA in the iSCNT embryos was found in any of the observed developmental stages (1- through 8-cell stages). In conclusion, bovine cytoplasm supports domestic cat nucleus development through the 8-cell stage. The mtDNA genotype of domestic cat-bovine iSCNT embryos illustrates persistence of heteroplasmy, and the reduction in mtDNA content might reflect a developmental block at the 8-cell stage.  相似文献   

15.
Effects of recipient oocyte activation methods on the development of nuclear transfer (NT) embryos were investigated. In Exp. 1, cell-cycle phase of serum-starved bovine cumulus cells was examined by flow cytometry. Majority (95.5%) of medium-sized (16-20 microm) cells that made up 56% of total cells was at the G0/G1 phase. NT embryos were constructed by electric fusion with the medium-sized serum-starved cumulus cells and bovine oocytes of 3 different preparations: enucleated oocytes treated with calcium ionophore A 23187 for 5 min and cycloheximide for 5 hr (A 23187/CHX), those treated with ethanol for 7 min and cycloheximide for 2 hr(ethanol/CHX) and those without treatment. In Exp. 2 and 3, developmental competence of NT embryos constructed with A 23187/CHX- and ethanol/CHX-treated oocytes was compared to that of NT embryos constructed with non-treated oocytes, respectively. Further, nuclear behavior in 3 different NT embryos was examined in Exp. 4. Within 1 hr after fusion, majority of the NT embryos constructed with non-treated oocytes showed condensed chromosome. Three hours after fusion, about 50% of NT embryos constructed with non-treated or ethanol/CHX-treated oocytes showed a single pronucleus-like structure. NT embryos constructed with ethanol/CHX-treated oocytes showed similar rates of fusion, cleavage and blastocyst formation to those of the non-treated oocytes. In contrast, NT embryos constructed with A 23187/CHX-treated oocytes did not show any pronucleus-like structure and showed lower cleavage rate and no development to blastocysts. The results indicate that ethanol/CHX-treated oocytes could support development of somatic cell NT embryos to the blastocyst stage at a similar rate to that of non-treated oocytes.  相似文献   

16.
以CZB为基础培养液,培养小鼠4、8-细胞胚胎单卵裂球,研究葡萄糖、牛磺酸和猪输卵管上皮细胞共培养在其体外发育中的作用。结果表明:牛磺酸添加与否对4、8-细胞胚胎单卵裂球的囊胚发育率分别为29%、30%和14%、14%,无显著性差异(P>0.05)。添加葡萄糖后,4、8-细胞胚胎单卵裂球的囊胚发育率分别为36%和19%,有显著提高(P<0.05)。含有葡萄糖而牛磺酸的添加与否对4、8-细胞胚胎单卵裂球的囊胚发育率分别为36%、38%和19%、20%,无显著性差异(P>0.05)。各组内4、8-细胞胚胎单卵裂球形成的囊胚细胞数分别为(10.44±1.24~(12.43±1.18)和(7.57±0.97)~(8.48±1.16),均无显著性差异(P>0.05)。4、8-细胞胚胎单卵裂球与猪输卵管上皮细胞共培养,其囊胚发育率分别为47%和26%,囊胚细胞数分别为(17.57±1.13)和(11.43±0.92),均高于单一培养(P<0.05)。  相似文献   

17.
The objective of this study was to evaluate the in vitro development of bovine embryos encapsulated in alginate. Day-4 embryos produced in vitro (n = 110) were encapsulated with 1.5% sodium alginate and co-cultured with oviduct cells. Unencapsulated embryos (n = 106) were used as controls. In vitro development rate to the blastocyst stage at Day 7 was similar between encapsulated, 42.7%, (47/110) and control. 34% (36/106). embryos. Although encapsulated embryos were able to hatch on Day 9, they did so in a lower proportion than controls (P < 0.05). In conclusion, alginate encapsulation of bovine embryos does not disturb the in vitro development up to the blastocyst stage but significantly reduces the hatching process.  相似文献   

18.
This study was conducted to reconstruct heterogeneous embryos using equine skin fibroblast cells as donor karyoplasts and the bovine oocytes as recipient cytoplast for investigating the reprogramming of equine somatic cell nuclear in bovine oocyte cytoplasm and the developmental potential of the reconstructed embryos. Adult horse skin fibroblast cells serum-starved were used as donor somatic cells. Bovine oocytes matured in vitro were employed as recipient cytoplasts. The fusion of fibroblast cells into recipient cytoplasm was induced by electofusion. The fused eggs were activated by inomycin with 2 mm/ml 6-dimethylaminopurine (6-DMAP). The activated reconstructed embryos were co-cultured with bovine cumulus cells in synthetic oviduct fluid supplemented with amino acid (SOFaa) and 10% fetal calf serum (FCS) for 168 h. The results showed that the first completed cleavage of xenonuclear transfer equine embryos occurred between 30 and 48 h following activation. 52% of the injected oocytes were successfully fused, 72% of the fused eggs underwent the first egg cleavage and 17% of the heterospecific nuclear-transferred zygotes developed to 4- or 8-cell embryo stages. This study demonstrated that the reconstructed embryos have undergone the first embryonic division and the reprogramming of equine fibroblast nuclei can be initiated in bovine-enucleated oocytes.  相似文献   

19.
The ability of frozen-thawed fetal skin was examined to generate viable cell lines for nuclear transfer. Fetal skin frozen at -35 degrees C or -80 degrees C in the presence of 5% DMSO were used as tissue explants to generate somatic cells. The resultant confluent cells were then used as donors for nuclear transfer (NT). Of the bovine NT embryos reconstructed from the somatic cells, 78% to 81% showed cleavage, 43% to 48% reached the stage of morula formation and 34% to 35% reached blastocyst formation. There were no significant differences in development (P>0.05) when the NT embryos were compared with those reconstructed from fresh somatic-cell-derived skin tissues (75%, 45%, and 38%, for cleavage, and development to morula and blastocyst stages, respectively). The results indicated that cell lines derived from bovine fetal skin cryopreserved by a simple method could be used as donors in nuclear transfer. The resulting embryos showed similar development in vitro to those reconstructed from unfrozen fetal-skin-derived somatic cells.  相似文献   

20.
影响小鼠早期胚胎发育的几个重要因素   总被引:2,自引:0,他引:2  
以Whiten氏液为基本培养液,研究了卵丘细胞、谷氨酰胺及不同浓度的葡萄糖对HCG注射后17~18h、25~27h和36~38h昆明小鼠胚胎体外发育的影响。结果表明,不论葡萄糖或谷氨酰胺存在与否,卵丘细胞与受精卵的共同培养对胚胎克服体外发育阻断无明显效果。葡萄糖能抑制1-细胞胚胎发育,促进4-细胞以后的胚胎发育。谷氨酰胺是小鼠早期胚胎发育的良好能源物质,可使96%(308/321)的体外发育到2-细胞的胚胎克服阻断而继续发育到4-细胞。谷氨酰胺与葡萄糖联合使用,则可使小鼠受精卵的囊胚发育率达57.5%(229/398)。HCG注射后36~38h的小鼠胚胎能利用葡萄糖。  相似文献   

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