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1.
试验旨在研究程序化冷冻前后小鼠正常孵化囊胚和休眠胚胎中C3蛋白的分布及转录水平的差异表达情况,为阐明哺乳动物胚胎在抗冻过程中的相关调控机制提供理论依据。试验选用ICR系雌性小鼠,从妊娠第5天小鼠子宫回收正常孵化囊胚;构建延迟着床模型获取小鼠休眠胚胎,利用激光共聚焦和实时荧光定量PCR技术对各组胚胎进行细胞免疫荧光和C3 mRNA相对表达量的检测。结果发现,程序化冷冻前后的正常孵化囊胚和休眠胚胎中C3在转录和翻译水平均有表达;正常孵化囊胚经冷冻处理后C3 mRNA相对表达量显著上调（P < 0.05）;休眠胚胎冷冻后C3 mRNA相对表达量与冷冻前相比显著下调（P < 0.05）;冷冻前休眠胚胎C3 mRNA相对表达量显著高于冷冻前正常孵化囊胚（P < 0.05）;冷冻后休眠胚胎C3 mRNA相对表达量显著低于冷冻后正常孵化囊胚（P < 0.05）。结果表明,胚胎中C3基因在转录水平的下调表达对胚胎抗冻具有一定的积极作用,C3基因很可能参与了胚胎抗冻过程的相关调控。 相似文献
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Yan CL Yang QE Zhou GB Hou YP Zhao XM Fan ZQ Liu MQ Liu L Zhu SE 《Acta veterinaria Hungarica》2008,56(2):245-253
The present study was designed to investigate the cryotolerance of in vitro fertilised (IVF) mouse embryos at various preimplantation developmental stages. IVF mouse embryos were vitrified by the open-pulled straw (OPS) method. After warming, embryos were morphologically evaluated and assessed by their development to blastocysts, hatched blastocysts or term. The results showed that a high proportion (93.3-100.0%) of vitrified embryos at all developmental stages were morphologically normal after recovery. The developmental rate of vitrified 1-cell embryos to blastocyst (40.0%) or hatched blastocyst (32.7%) or term (9.3%) was significantly lower than that from other stages (P < 0.05). Vitrified embryos from 2-cell to early blastocyst stage showed similar blastocyst (71.8-89.5%) and hatched blastocyst rates (61.1-69.6%) and could develop to term without a significant loss of survival compared with those of fresh embryos (P > 0.05). Vitrified 2-cell embryos showed the highest survival rate in vivo (50.6%, 88/174), compared with that from other stages (9.3-30.5%, P < 0.05). The data demonstrate that the OPS method is suitable for the cryopreservation of IVF mouse embryos from 2-cell stage to early blastocyst stage without a significant loss of survival. Embryos at the 2-cell stage had the best tolerance for cryopreservation in the present study. 相似文献
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旨在探讨初次卵裂时间对猪孤雌胚胎发育潜能及其基因相对表达水平的影响。本试验从健康母猪卵巢上抽取卵母细胞进行体外成熟培养，将猪孤雌激活胚胎分为早期卵裂组（16~22 h）与晚期卵裂组（26~32 h）统计比较卵裂率和囊胚率，并对囊胚的多能性相关基因Oct4、Sox2、Klf4等和凋亡相关基因Bcl-xl、Bax、Caspase-3的相对表达水平进行分析检测。结果表明，猪孤雌激活胚胎在16~22 h发生卵裂的为50%~60%，而26 h之后发生卵裂的不到20%，在18 h前完成第一次卵裂的胚胎囊胚发育率为79%，42 h后发生初次卵裂的胚胎无法发育至囊胚期。猪孤雌激活胚胎早期卵裂组的囊胚发育率显著高于晚期卵裂组（P<0.05）。早期卵裂组囊胚的Oct4、Nanog、Sox2、Klf4基因的表达量显著高于晚期卵裂组（P<0.05），Oct4、Sox2、Klf4基因的相对表达量极显著高于晚期卵裂组（P<0.01），Bax和Caspase-3基因的相对表达水平极显著低于晚期卵裂组（P<0.01），而Bcl-xl作为保护因子其表达量相对于晚期卵裂胚胎（26~32 h）显著上调（P<0.05）。结果显示，初次卵裂时间较早的猪孤雌激活胚胎发育潜能显著高于较晚卵裂胚胎，其囊胚多能性相关基因表达上调，凋亡相关基因表达下调，卵裂时间可作为鉴定猪孤雌激活胚胎发育潜力的重要参数。 相似文献
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Study on Time Period Model for Efficiently Collecting Hatched Blastocysts Flushed from Murine Uterus
ZHAI Chun-dong GU Mei-chao LIU Di WANG Li-hong QI Xiao-long ZHANG Jia-nan LIU Yun-hai NI He-min GUO Yong 《中国畜牧兽医》2015,42(5):1253-1258
The persent study was aimed to investigate the optimization of mice cage time after superovulation, and efficient acquisition of mouse hatched blastocysts.The experiment selected 150 ICR female mice aged 8 weeks, were randomly divided into 5 groups, the same time superovulation treatment according to 1:1, after the male and female in 18:00, 19:00, 20:00, 21:00 alloy cage overnight, the next day as early as 08:00 check, found that vaginal suppository for the first day of the pregnancy (D1).Take the pregnant the fifth day (D5) mice were sacrificed, their bilateral uterine horns, rushed from the embryonic.Statistics each thrust ratio and the total number of embryos hatched blastocysts/take not hatched blastocysts, was used as the index to evaluate embryos to obtain efficiency;Statistical trophectoderm cell number/inner cell mass number, as a reference index to evaluate the quality of embryo.The results found that groups of Ⅰ, Ⅱ and Ⅲ were no significant difference in the blastocyst number (P>0.05), but there was increasing trend, group Ⅳ was significantly higher than the other three groups (P<0.05).Groups Ⅰ, Ⅱ, Ⅲ and Ⅳ within the cell mass cells number/trophectoderm cell number were 23.18%, 23.55%, 21.72% and 23.28%, there was no significant difference in each group (P>0.05).The results showed that the corresponding set of Ⅳ cage got the time of mouse hatched blastocysts to obtain the highest efficiency, there was no significant differences in embryonic blastocyst quality. 相似文献
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本试验旨在优化小鼠超排后的合笼时间,高效获取小鼠孵化囊胚。试验选用150只ICR系8周龄雌性小鼠,随机分为5组,同一时间超排处理后雌、雄按1:1于18:00、19:00、20:00、21:00合笼过夜,次日上午08:00查栓,发现阴道栓这为妊娠第1天(D1)。取妊娠第5天(D5)小鼠处死,剪取双侧子宫角,冲取胚胎。统计每组冲取胚胎的总数及孵化囊胚/未孵化囊胚的比值,作为胚胎获取效率的评价指标;统计内细胞团数/滋养外胚层细胞团数,作为评价胚胎质量的参考指标。结果发现,在数量上组Ⅰ、组Ⅱ、组Ⅲ囊胚数差异不显著(P>0.05),但有增高趋势,组Ⅳ囊胚数显著高于其他3组(P<0.05)。组Ⅰ、组Ⅱ、组Ⅲ、组Ⅳ内细胞团数/滋养外胚层细胞团数分别为23.18%、23.55%、21.72%和23.28%,各组间差异不显著(P>0.05)。结果表明,组Ⅳ所对应的合笼时间获取小鼠孵化囊胚获取效率最高,胚胎囊胚质量无明显差异。 相似文献
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Conventional freezing of in vitro-produced and biopsied bovine blastocysts in the presence of a low concentration of glycerol and sucrose 总被引：1，自引：0，他引：1
The purpose of this study was to develop a practical cryopreservation method for in vitro-produced (IVP) and sex-predetermined bovine blastocysts that will be applicable to direct transfer of the post-thaw embryos. Blastocysts were harvested 7 days after IVF and allocated to either an intact or biopsy group. The cryoprotective solution contained 0.7 M glycerol and 0, 0.05 or 0.1 M sucrose. Slow cooling at a rate of -0.5 C/min was terminated at -25, -30, or -35 C, and rapid cooling in liquid nitrogen was followed. After one-step thawing and dilution, the IVP blastocysts were cultured for 3 days to assess their survival. The post-thaw survival rate of intact blastocysts after termination of slow cooling at -30 C in 0.7 M glycerol plus 0.1 M sucrose (96.2%) was significantly higher than that at -25 C in 0.7 M glycerol alone (44.4%). The post-thaw survival rate of biopsied bovine blastocysts after termination of slow cooling at -25 C in 0.7 M glycerol alone (53.8%) tended to be lower than that at -25 C in 0.7 M glycerol plus 0.05 M sucrose (91.3%) or -30 C in 0.7 M glycerol plus 0.1 M sucrose (92.3%). Thus, addition of a small amount of sucrose to 0.7 M glycerol cryoprotective solution shortened the process of slow cooling for both the intact and biopsied bovine embryos. Judged from the survival levels in vitro after thawing and one-step dilution of embryos (>80%), this is an improved method of cryopreservation for subsequent direct transfer of IVP and biopsied bovine blastocysts. 相似文献
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山羊类ES细胞的分离与克隆 总被引：6，自引：0，他引：6
采集山羊交配后6～8d的桑椹胚、囊胚和孵化囊胚,将桑椹胚和囊胚分别放在小鼠原代胎儿成纤维细胞(PMEF)饲养层和同源原代胎儿成纤维细胞(PGEF)饲养层上比较其脱带时间及脱带率。脱带后,将各自一半胚胎切割,把含ICM的半胚分别放在相应饲养层上进行培养,另一半整胚在各自饲养层上继续培养,而孵化囊胚直接于PGEF饲养层上培养。当ICM增殖一定程度时进行传代,以比较其类ES细胞分离与克隆的效果。结果表明,在2种不同饲养层上,囊胚的脱带时间均短于桑椹胚,囊胚的脱带率均高于桑椹胚,而饲养层的种类对胚胎的脱带时间以及脱带率影响不大。脱带切割囊胚不论在PMEF还是在PGEF饲养层上,其贴壁时间均短于脱带整胚及孵化囊胚,而贴壁率高于脱带整胚,与孵化囊胚相似。脱带整胚及脱带切割胚在PMEF饲养层上所获类ES细胞只能维持3代,而在PGEF饲养层上,脱带切割半胚和孵化囊胚所获类ES细胞传至5代。由此认为,对脱带后的胚胎进行切割处理,有利于ICM的贴壁和增殖;应用同源原代胎儿成纤维细胞饲养层培养系统,有利于类ES细胞的分离与克隆。 相似文献
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小鼠2-细胞胚胎细管法和OPS法玻璃化冷冻保存技术的研究 总被引：8，自引：0，他引：8
本试验在室温 (2 0℃和 2 5℃ )条件下 ,利用不同浓度的玻璃化溶液 (EFS和EDFS) ,对小鼠 2 细胞胚胎进行细管法和OPS法玻璃化冷冻保存。在 2 0℃室温条件下 ,用EFS4 0平衡 1min细管一步法冷冻 ,解冻后囊胚发育率仅为35 .0 % ,和新鲜 2 细胞体外培养的对照组 (6 5 .0 % )的差异极显著 (P <0 .0 1)。当 2 细胞胚胎在 10 %EG +10 %D溶液中预处理 5min ,再移入EDFS中平衡 30s二步法冷冻保存 ,解冻后囊胚发育率达 4 7.8%～ 4 8.8% ;当室温升至2 5℃时 ,二步法冷冻保存后 2 细胞的囊胚发育率达到 5 2 .2 % ,与对照组无显著差异 (P >0 .0 5 )。改用OPS二步法EFS30冷冻组保存后的 2 细胞胚胎的囊胚发育率高达 6 2 .2 % ,为试验中的最佳组。用最佳细管法和OPS法冷冻组解冻后培养至囊胚移植给受体母鼠均获得产仔 相似文献
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探讨程序化冷冻与玻璃化冷冻对小鼠GV期卵母细胞及二细胞期胚胎的复苏率及其发育潜能的影响。通过小鼠的卵母细胞与早期胚胎的不同冷冻方法的比较,为后续阿旺绵羊的胚胎冷冻保存提供参考。采用程序化冷冻与玻璃化冷冻技术,分别冷冻小鼠GV期卵母细胞及二细胞期胚胎,复苏后培养,比较不同冷冻处理后的复苏率、成熟率与囊胚率。小鼠GV期卵母细胞程序化冷冻复苏率(48.00%±5.29%)显著低于玻璃化冷冻复苏率(65.00%±5.00%),有统计学差异(P=0.0147<0.05);而程序化冷冻后复苏卵母细胞的发育成熟率略高于玻璃化冷冻组,但无统计学意义。小鼠二细胞期胚胎程序化冷冻组复苏率(76.00%±2.00%)显著高于玻璃化冷冻组复苏率(70.00%±2.00%),有统计学差异(P=0.0213<0.05);冷冻后复苏胚胎发育的囊胚率程序化冷冻略低于玻璃化冷冻及对照组,但无统计学意义。 相似文献
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内皮素3对小鼠黑色素沉着相关基因表达的影响 总被引：1，自引：1，他引：0
试验旨在探究内皮素3（endothelin 3，EDN3）在不同毛囊时期小鼠皮肤中是否存在差异表达及其对黑色素细胞黑色素沉着相关基因表达的影响。通过RT-PCR技术对EDN3、EDNRB在不同毛囊时期小鼠皮肤的表达情况进行定量分析发现，EDN3和EDNRB在小鼠不同毛囊时期皮肤样品中均有表达，在毛囊生长初期和中期小鼠皮肤中EDN3 mRNA表达量是末期的2.18倍（P < 0.05）和1.15倍（P > 0.05），毛囊生长初期和中期EDNRB mRNA表达量是末期的16.8倍（P < 0.01）和9.9倍（P < 0.01）。为了进一步揭示EDN3在黑色素细胞色素沉着中的重要作用，通过细胞转染技术使小鼠黑色素细胞过表达EDN3并测定其黑色素含量及色素沉着相关基因的表达水平发现，与空载组相比，体外转染EDN3的黑色素细胞黑色素含量明显增加。此外，MITF mRNA显著降低（P < 0.05）；TYR、TYRP2、EDNRB、c-Kit和EDN1 mRNA显著升高2.04倍（P < 0.05）、1.44倍（P < 0.05）、1.41倍（P < 0.05）、5.21倍（P < 0.01）和3.27倍（P < 0.01）。MITF蛋白水平显著降低（P < 0.05），TYR、TYRP2、EDNRB和c-Kit蛋白水平显著升高1.48倍（P < 0.01）、4.61倍（P < 0.01）、1.27倍（P < 0.05）和2.64倍（P < 0.01）。综上所述，EDN3在不同毛囊时期小鼠皮肤中均可有效表达，且表达量存在显著差异。黑色素细胞中过量表达EDN3，使色素沉着相关基因的表达量及黑色素含量增加。 相似文献
13.
Miyuki MORI Takeshi HAYASHI Yoshihiro ISOZAKI Naoki TAKENOUCHI Miki SAKATANI 《The Journal of reproduction and development》2015,61(5):423-429
In this study, the effect of heat shock on frozen-thawed blastocysts was evaluated using in
vitro-produced (IVP) bovine embryos. In experiment 1, the effects of 6 h of heat shock at 41.0 C on
fresh blastocysts were evaluated. HSPA1A expression as a reflection of stress was increased
by heat shock (P < 0.05), but the expressions of the quality markers IFNT and
POU5F1 were not affected. In experiment 2, frozen-thawed blastocysts were incubated at 38.5
C for 6 h (cryo-con) or exposed to heat shock at 41.0 C for 6 h (cryo-HS). Then, blastocysts were cultured at
38.5 C until 48 h after thawing (both conditions). Cryo-HS blastocysts exhibited a decreased recovery rate:
HSPA1A expression was dramatically increased compared with that in fresh or cryo-con
blastocysts at 6 h, and IFNT expression was decreased compared with that in cryo-con
blastocysts at 6 h (both P < 0.05). Cryo-con blastocysts at 6 h also exhibited higher
HSPA1A expression than fresh blastocysts (P < 0.05). At 48 h after thawing, the number
of hatched blastocysts and blastocyst diameter were lower in cryo-HS blastocysts (P < 0.05). Cryo-con
blastocysts showed lower POU5F1 levels at 48 h than fresh, cryo-con or cryo-HS blastocysts at
6 h (P < 0.05), but their POU5F1 levels were not different from those of cryo-HS
blastocysts at 48 h. These results indicated that application of heat shock to frozen-thawed blastocysts was
highly damaging. The increase in damage by the interaction of freezing-thawing and heat shock might be one
reason for the low conception rate in frozen-thawed embryo transfer in summer. 相似文献
14.
本实验旨在探讨昆明白小鼠桑椹胚超低温冷冻后Oct-4表达的变化与胚胎发育潜力的关系。胚胎采用OPS法冷冻,即胚胎于10%EG+10%DMSO溶液中预处理30 s,然后再移入玻璃化溶液(EDFS30)中处理25 s,以OPS为承载器投入液氮中。毒性组胚胎未投入液氮,其他过程与冷冻组相同,新鲜胚胎为对照组。胚胎解冻后,检测Oct-4 mRNA与蛋白的表达,通过观察其形态正常率、囊胚发育率和囊胚细胞数来综合判断其体外发育能力。结果表明:冷冻组和毒性组较桑椹胚中Oct-4 mRNA的表达量对照组相比显著降低(P<0.05),蛋白表达量3组间无显著差异。桑椹胚处理后的形态正常率以及桑椹胚培养48 h后的囊胚发育率(96.67%～100%)和囊胚细胞数(89.67～92.33)3组间无显著差异。结果显示,玻璃化冷冻保存小鼠桑椹胚后多能性基因Oct-4 mRNA表达的变化并未影响其体外发育潜力。 相似文献
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Cloned porcine embryos can maintain developmental ability after cryopreservation at the morula stage
Nakano K Matsunari H Nakayama N Ogawa B Kurome M Takahashi M Matsumoto M Murakami H Kaji Y Nagashima H 《The Journal of reproduction and development》2011,57(2):312-316
The aim of the present study was to clarify the overall efficiency of porcine somatic cell nuclear transfer (SCNT) by incorporating cryopreservation of the cloned embryos before transfer. The SCNT embryos reconstructed with preadipocytes and in vitro-matured (IVM) oocytes were cultured to harvest morula stage embryos; they were then subjected to delipation (removal of cytoplasmic lipid droplets) and vitrification. After warming and culture, the embryos developing to blastocysts were transferred to recipients to obtain cloned piglets. From 372 reconstructed embryos, 188 (50.5%) reached the morula stage and 117 (31.5%) developed to blastocysts after vitrification. Transfer of 98 (26.3%) morphologically normal blastocysts gave rise to 6 (1.6%) piglets, including 1 stillborn. The efficiency of the cloned piglet production was comparable with that obtained using SCNT embryos without cryopreservation (2.7%, 17/635). Here, we demonstrate that porcine somatic cell cloning can be performed without a significant reduction in efficiency even when the SCNT embryos are cryopreserved before transfer. 相似文献
16.
Hochi S 《The Journal of reproduction and development》2003,49(1):13-21
Factors affecting sensitivity of preimplantation embryos and follicular oocytes to cryopreservation were analyzed in the equine and bovine species. (1) Survival of equine blastocysts after two-step freezing in the presence of glycerol as the cryoprotective agent (CPA) was influenced by development of the embryonic capsule. The use of ethylene glycol (EG) with sucrose as CPAs improved the post-thaw survival of blastocysts and made it possible to transfer the embryos into recipient mares without removing the CPAs. In addition, early blastocysts cryopreserved by vitrification could develop both in vitro and in vivo when the embryos were exposed to vitrification solution in a stepwise manner. The vitrification procedure was also applied to the relatively large expanded blastocysts. (2) Bovine embryos produced in vitro have been considered to be highly sensitive to the process of cryopreservation. To solve this problem, Day-7 blastocysts produced in a serum-free system were cooled at 0.3 C/min rather than 0.6 C/min before being plunged into liquid nitrogen, resulting in no loss of the post-thaw viability. The supplementation of LAA in IVM/IVF media or IVC medium was effective in producing pronuclear-stage zygotes or morula-stage embryos relatively tolerable to freezing, respectively. (3) Transmission electron microscopic observation of immature equine oocytes showed that cellular injury occurred near the sites of gap-junctions between cumulus cells and the oocyte. In cattle, higher fertilization rates of oocytes were obtained when the oocytes were subjected to cryopreservation at an intermediate stage during IVM (GVBD for freezing, Met-I for vitrification). Vitrification of bovine Met-II oocytes in open-pulled glass capillaries, characterized by an ultra-rapid cooling rate (3,000-5,000 C/min), was found to avoid any harmful influence of vitrification and warming. 相似文献
17.
【目的】研究柠檬苦素(limonin,Lim)对小鼠卵母细胞体外成熟(IVM)及后续体外受精(IVF)胚胎发育潜能的影响,旨在为体外成熟培养系统的优化提供参考。【方法】在小鼠体外成熟培养液中添加不同浓度的Lim(0、10、20、50 μmol/L),成熟培养12 h后统计小鼠卵母细胞第一极体(PBI)排出率,筛选体外成熟培养液中添加Lim的最适浓度;在体外成熟培养液中添加最适浓度的Lim,以0 μmol/L Lim为对照组,成熟培养12 h,通过免疫荧光染色检测活性氧(ROS)、谷胱甘肽(GSH)以及线粒体膜电位(MMP)水平;通过实时荧光定量PCR检测卵母细胞抗氧化及凋亡相关基因的mRNA表达水平。将最适Lim组及对照组卵母细胞体外成熟24 h后进行体外受精,于体外受精24 h和3.5 d分别统计胚胎卵裂率和囊胚率,并用Fluorescein-dUTP和Hoechst 33342染色分别检测囊胚总细胞数及囊胚内凋亡细胞比率。【结果】与0 μmol/L Lim组相比,20 μmol/L Lim组小鼠卵母细胞PBI排出率显著升高(P＜0.05),后续试验均用20 μmol/L Lim进行处理。与对照组组相比,20 μmol/L Lim组小鼠卵母细胞内ROS水平显著降低(P＜0.05),GSH、MMP水平均显著增加(P＜0.05),抗氧化相关基因(GPx3、CAT和Prdx3)、抗凋亡相关基因(Bcl-2、Bcl-xl)表达水平均显著上调(P＜0.05),促凋亡相关基因(Caspase-3)表达水平显著下调(P＜0.05);体外受精胚胎的卵裂率、囊胚率、囊胚总细胞数均显著增加(P＜0.05),囊胚内细胞凋亡比率显著下降(P＜0.05)。【结论】在体外成熟培养液中添加20 μmol/L Lim可以通过抑制氧化应激和细胞凋亡、增加MMP水平提高小鼠卵母细胞质量,从而提高体外受精胚胎的发育潜力。 相似文献
18.
Solid surface vitrification (SSV) was compared with in-straw vitrification for cryopreservation of biopsied mouse embryos. Eight-cell stage embryos were zona drilled and one blastomere was removed. Developed morulae or blastocysts were vitrified in microdrop (35% EG + 5% PVP + 0.4 M trehalose) or in straw (7.0 M EG + 0.5 M sucrose). Following recovery, embryos were cultivated in vitro or transferred into recipients. Cryopreservation had an effect not only on the survival of biopsied embryos but also on their subsequent development in vitro. Cryosurvival of biopsied morulae vitrified in straw was significantly inferior to SSV. The post-warm development of biopsied and non-biopsied morulae was delayed on Day 3.5 and 4.5 in both vitrification groups. A delay in development was observed on Day 5.5 among vitrified non-biopsied blastocysts. The percentage of pups born from biopsied morulae or blastocysts following cryopreservation did not differ from that of the control. No significant differences could be detected between methods within and between embryonic stages in terms of birth rate. The birth rate of biopsied embryos vitrified in straw was significantly lower compared to the non-biopsied embryos. The novel cryopreservation protocol of SSV proved to be effective for cryopreservation of morula- and blastocyst-stage biopsied embryos. 相似文献
19.
通过对不同发育阶段猪孤雌激活胚胎进行离心去脂处理，比较胚胎冷冻效果，优化冷冻体系。以猪孤雌激活胚胎为材料，比较实施电激活后第1天（2～3细胞）、第2天（4～6细胞）、第3天（6～8细胞）进行离心处理，取培养到第6天所得到扩张囊胚进行玻璃化冷冻保存，未离心处理直接进行冷冻的胚胎作为对照组。结果显示第3天离心组和第2天离心组所得的囊胚形成率（30．23％，28．22％）虽好于第1天离心组和不处理组（21．73％，22．43％），但4个试验组差异不显著（P〉0．05）；第3天离心组和第2天离心组的冷冻后胚胎复苏率（28．57％，20．56％）显著高于不处理组（7．73％）、第1天离心组（12．24％），差异显著（P〈0．05）；第3天离心组的解冻后囊胚内细胞计数（30．40）显著高于对照组（18．25）、第1天离心组（16．60），差异显著（P〈0．05），高于第2天处理组（23．71），但差异不显著（P〉0．05）。结果表明，在孤雌激活后第3天进行离心处理，对胚胎发育到囊胚没有影响，且进行冷冻后所得复苏率及胚胎内细胞团数较好。 相似文献
20.
鹌鹑早期胚胎bcl-2基因表达的差异及发育性变化 总被引：1，自引：1，他引：0
采用RT-PCR方法,用Wpkci和β-actin引物进行多重PCR鉴定66～120 h鹌鹑胚胎样本性别后,选取不同时间点雌、雄胚胎各4枚,以β-actin为内标,测定bcl-2 mRNA的相对丰度;探讨bcl-2对鹌鹑早期胚胎发育的影响。结果表明：①雄性胚胎在66～90 h bcl-2 mRNA表达量一直维持在较低水平,96～102 h bcl-2 mRNA有所增加,但96与102 h间差异不显著（P＞0.05）,在108 h显著降低（P＜0.05）;与108 h相比,114 h显著升高,120 h维持在较高水平。雄性胚胎在66～108 h（72 h除外）bcl-2 mRNA表达均比雌性胚胎的表达量低,在114～120 h bcl-2 mRNA表达水平均比雌性胚胎的高。②雌性胚胎在66～90 h bcl-2 mRNA表达量一直维持在较低水平;与90 h相比,96 h显著升高,96～120 h逐渐降低,其中96～108 h下降,但三者间差异不显著（P＞0.05）;与108 h相比,114～120 h显著下降（P＜0.05）。③相同时间点雌、雄胚胎间的比较结果表明,84 h雌性胚胎bcl-2 mRNA表达显著高于雄性胚胎（P＜0.05）,90、108 h雌性胚胎bcl-2 mRNA表达极显著高于雄性胚胎（P＜0.01）;而在120 h雄性胚胎bcl-2 mRNA表达极显著高于雌性胚胎（P＜0.01）。鹌鹑胚胎发育早期,雌、雄样本bcl-2 mRNA表达存在一定的时序性,说明bcl-2对雌、雄鹌鹑早期分化及早期胚胎发育有重要影响。 相似文献