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1.
目的:研究表皮生长因子(epidermalgrowthfactor,EGF)在小鼠早期胚胎体外发育中的作用,以及EGF联合胰岛素样生长因子-Ⅱ(insulin-likegrowthfactor-II,IGF-Ⅱ)对小鼠早期胚胎体外发育是否有协同促进作用。方法:①在mKSOM培养液中添加0ng/ml(对照组)、0.1ng/ml、1ng/ml、10ng/ml、100ng/ml的EGF、10ng/mlIGF-Ⅱ、1ng/mlEGF+1ng/mlIGF-Ⅱ、1ng/mlEGF+10ng/mlIGF-Ⅱ、10ng/mlEGF+1ng/mlIGF-Ⅱ、10ng/mlEGF+10ng/mlIGF-Ⅱ培养小鼠1-细胞胚胎,每组胚胎在培养箱中连续培养120h,每24h观察胚胎发育情况,分别计算2-细胞率、4-细胞率、桑椹胚率、囊胚率和孵化率,并进行囊胚细胞计数。结果:添加1ng/ml、10ng/mlEGF组的囊胚率及孵化率显著增加;EGF+IGF-Ⅱ组较单一添加组、对照组囊胚率及孵化率显著增加(P<0.05),且EGF+IGF-Ⅱ组囊胚细胞计数显著高于其他组(P<0.05),其中10ng/mlEGF+10ng/mlIGF-Ⅱ组的孵化率及囊胚细胞数最高(P<0.05)。结论:EGF浓度在1 ̄10ng/ml范围内可以提高体外培养鼠胚的囊胚率及孵化率;EGF及IGF-Ⅱ对体外培养鼠胚的发育有协同促进作用;本实验范围内10ng/mlEGF+10ng/mlIGF-Ⅱ为最优组合。  相似文献   

2.
朱伟杰  李菁  张文红  姚康寿 《生殖与避孕》2004,24(1):19-23,T004
目的:探讨人精子培养试验检测培养液内毒素的敏感性。方法:活动精子分别与6个内毒素浓度(0.5 ng/mL,1 ng/mL,10 ng/mL,1 000 ng/mL,10 000 ng/mL,50 000 ng/mL;n=10)共培养,检测培养后的精子活动率,比较实验组与对照组的精子活动率改变,并观察不含白蛋白培养液的3种内毒素浓度(1 000 ng/mL,10 000 ng/mL,50 000 ng/mL;n=6)对精子活动率的影响。另对比小鼠1-细胞胚胎和2-细胞胚胎培养试验对内毒素(0.5 ng/mL,1 ng/mL,10 ng/mL)的敏感性。结果:精子培养24 h后,在含白蛋白的内毒素浓度为0.5-1 000 ng/mL的4组中,实验组的精子活动率与对照组的比较没有显著性差异(P>0.05),但在内毒素浓度为10 000 ng/mL和50000ng/mL组,精子活动率则显著降低(P<0.01)。在不含白蛋白的内毒素浓度为50 000 ng/mL组和10 000 ng/mL组中,培养2 h和8 h的精子活动率分别显著降低(P<0.01)。在培养液内毒素浓度为0.5 ng/mL组和1 ng/mL组,小鼠1-细胞胚胎和2-细胞胚胎各发育阶段的发育率均显著减少,内毒素对胚胎发育的抑制呈剂量效应。结论:人精子存活试验可检测培养液中较高的内毒素水平,其敏感性比小鼠1-细胞胚胎和2-细胞胚胎培养试验的低。培养液中不含白蛋白可提高精子对内毒素的敏感性。  相似文献   

3.
目的:研究CD82/KAI1mRNA及蛋白质在小鼠胚胎中的表达规律及其对小鼠胚胎体外发育的影响。方法:①应用RT-PCR及免疫荧光技术观察CD82/KAI1mRNA及蛋白质在小鼠不同发育时期胚胎中的表达规律。②在不同CD82/KAI1抗体浓度的培养液中,培养小鼠8-细胞胚胎,观察囊胚发育率、孵化率和胚胎细胞数的变化。结果:①CD82/KAI1mRNA在小鼠不同时期胚胎中均有表达,于8-细胞期及桑葚胚表达较丰富,CD82/KAI1蛋白表达于各期胚胎细胞的胞膜和胞浆,在桑葚胚期CD82蛋白还强表达于胚胎细胞的胞核。②一定浓度的CD82/KAI1抗体可明显抑制胚胎的发育速度(1∶800,P<0.05),降低囊胚的形成率(1∶400,P<0.05)和孵出率(1∶800,P<0.05),减少囊胚细胞数(1∶800,P<0.05)。结论:CD82/KAI1在小鼠胚胎的发育过程中可能发挥重要作用。  相似文献   

4.
朱颖  吴超英  孙永玉 《生殖与避孕》2004,24(5):262-264,278
目的:探讨胎盘异铁蛋白对小鼠胚胎生长发育的影响。方法:将人早孕蜕膜细胞和小鼠2-细胞期胚胎置于含不同浓度胎盘异铁蛋白的培养液中共培养,倒置显微镜下观察记录胚胎数目并分级。结果:①发育至4-细胞期、8-细胞期及桑葚胚阶段鼠胚数的百分率比较,差异无显著性(P>0.05)。②在10 U/mL和100 U/mL浓度的胎盘异铁蛋白作用下,发育至囊胚期和孵出期鼠胚数的百分率明显提高(P<0.05)。③在1 000 U/mL浓度的胎盘异铁蛋白作用下,从2-细胞期发育至孵出期鼠胚数的百分率有所下降,部分细胞发生变性、退化和不规则分裂等现象,但无统计学意义(P>0.05)。结论:10~100 U/mL浓度的的胎盘异铁蛋白对鼠胚的早期发育没有显著影响,但能促进晚期鼠胚的生长、分化和孵出。  相似文献   

5.
张嬿  石红 《生殖与避孕》2003,23(6):323-326
目的:研究人子宫内膜共培养体系对早期鼠胚体外发育的影响及移植后的妊娠情况。方法:将2-细胞小鼠胚胎与人子宫内膜细胞进行体外共培养,对照组为无营养细胞的单纯培养液,每日在显微镜下观察胚胎的发育情况。将培养到囊胚期的胚胎移植回小鼠的子宫腔,观察着床情况。结果:共培养体系中68.3%的2-细胞胚胎发育至桑椹胚期,50.8%发育至囊胚期,囊胚的孵化率为36.7%,胚胎的着床率为25.0%。而对照组只有24.8%的2-细胞胚胎发育至桑椹胚期,11.4%到达囊胚期,且其中大部分为早期囊胚即停止发育。另外对照组细胞碎片出现早且多,卵裂球不均匀,胚形态差,移植后胚胎的着床率仅为3.1%。结论:人子宫内膜细胞共培养体系可以促进小鼠胚胎的体外发育,改善胚胎的质量,提高着床率。  相似文献   

6.
目的:探讨体外受精(IVF)周期中胚胎细胞数量和碎片比例对囊胚形成的影响。方法:回顾性分析2 404个IVF周期患者的13 037个进行囊胚培养的胚胎。根据第3日细胞数量将胚胎分为2~3-细胞组(2C~3C)、4-细胞组(4C)、5-细胞组(5C)、6-细胞组(6C)、7-细胞组(7C)、8-细胞组(8C)、9-细胞组(9C)、10-细胞组(10C)、11-细胞组(11C)、12-细胞组(12C)、≥13-细胞组(≥13C),分析囊胚培养细胞数量和碎片比例与囊胚形成的关系。结果:碎片为1~2级的胚胎,8C组和≥13C组囊胚形成率明显高于其他各组(P0.05);7C、9C~12C组之间囊胚形成率无统计学差异(P0.05)。碎片为3级的胚胎,8C组的囊胚形成率高于其他各组(P0.05),且与碎片1~2级7C、9C~12C组的胚胎相比无统计学差异(P0.05);9C组囊胚形成率高于7C组,且差异显著(P0.05)。结论:第3日8C胚胎的发育潜能是最优的,可适当放宽对8C碎片的评估;随着碎片比例增大可减弱胚胎的发育潜能。  相似文献   

7.
废弃胚胎继续囊胚培养研究   总被引:10,自引:0,他引:10  
目的:探讨体外受精治疗周期中废弃胚胎的体外发育潜能。方法:通过囊胚序贯培养法将无原核(0PN)、单个原核(1PN)、多个原核(≥3PN)和卵裂期发育延缓的2原核(2PN)废弃胚胎培养至囊胚期。比较不同来源胚胎囊胚形成情况和d3胚胎的卵裂球数、质量分级等;并利用废弃胚胎囊胚形成情况对体外受精妊娠结局进行预测。结果:共收集801个废弃胚胎,经序贯培养,形成209个囊胚(26.09%),其中58个为优质囊胚(27.75%)。1PN胚胎、0PN胚胎、d3卵裂球数为7-9-细胞胚胎、d3评分为Ⅰ-Ⅱ级胚胎以及卵裂球数为偶数的胚胎囊胚形成率较高(P均<0.05)。废弃胚胎中有囊胚形成者的临床妊娠率明显高于无囊胚形成者(P<0.05)。结论:废弃胚胎有不同程度的发育潜能,部分可发育为囊胚;特别是:①0PN和1PN胚胎;②d3的4-9-细胞胚胎,偶数卵裂球者更佳;③I级和Ⅱ级胚胎。  相似文献   

8.
目的:观察冷冻前人工皱缩小鼠扩张期囊胚的囊胚腔,冻融后小鼠扩张期囊胚的存活率及体外和体内继续发育能力。方法:冻前先应用自制的玻璃微细管人工皱缩小鼠囊胚腔后,再应用冷冻环进行玻璃化冷冻小鼠扩张期囊胚150个。冻融后培养3h观察胚胎存活情况,以新鲜扩张囊胚为对照,并将存活胚胎及新鲜未冻囊胚移植受体孕鼠子宫内,观察胚胎的妊娠率和产仔率。结果:人工皱缩后小鼠扩张囊胚的存活率及孵出率均显著高于未皱缩组(P<0.05),皱缩组与对照组无显著差异(P>0.05)。移植后皱缩组的妊娠率及产仔率均显著高于未皱缩组(P<0.05),皱缩组与对照组无显著差异(P>0.05)。结论:冻前人工皱缩小鼠扩张期囊胚的囊胚腔后,再应用冷冻环进行玻璃化冷冻是个行之有效的方法,能明显提高囊胚的玻璃化冷冻效率。  相似文献   

9.
目的:比较三种玻璃化方法冷冻小鼠胚胎的存活率及继续发育能力的差异。方法:将573个小鼠2-细胞期胚胎随机分为四组:Straw冷冻组144个,OPS冷冻组142个,CPS冷冻组149个,比较胚胎复苏后存活率、囊胚形成率及孵化率,并以未冷冻为对照组(138个)。结果:Straw组及CPS组存活率均显著性高于OPS 组(P <0.05);囊胚形成率及孵化率OPS组和CPS组均显著性高于Straw组(P均<0.05),OPS和CPS两组之间无显著性差异(P均>0.05)。结论:OPS与CPS方法能够更好地保持胚胎复苏后的继续发育能力。且CPS方法存活率高于OPS方法,并降低了交叉感染的危险性,更具临床应用潜力。  相似文献   

10.
人废弃胚胎单卵裂球体外发育潜能研究   总被引:4,自引:3,他引:1  
目的:探讨体外受精治疗周期废弃胚胎来源的单卵裂球体外发育潜能。方法:收集21例患者授精培养后第3日不适于移植和冷冻的废弃胚胎共60枚,其中2-细胞期胚胎10枚,3-细胞期胚胎8枚,4-细胞期胚胎26枚,5-细胞期胚胎12枚,6-细胞期胚胎4枚。采用化学法去除透明带并分割胚胎获得单卵裂球,继续体外培养,每24h观察卵裂球发育情况。结果:60枚胚胎共获得229枚单卵裂球。以分割取得单卵裂球日作为第1日(d1),继续培养,其发育似乎遵循d2分裂、d3紧密化、d4出现囊胚腔、d5扩张,延续分割前的程序,但提前进入紧密化期,致使囊胚细胞数少,囊胚期体积亦比正常胚胎小。其中,4-细胞期废弃胚胎来源的卵裂球分裂率、紧密化率、囊胚形成率显著高于来自其它细胞数的单卵裂球(P<0.05),且其大部分内细胞团(ICM)细胞的碱性磷酸酶(AKP)表达呈阳性。结论:废弃胚胎来源的单卵裂球能在体外继续发育至囊胚,发育潜能与供体胚胎发育质量密切相关,其发育似乎延续原有供体卵裂球的节律,而形成囊胚所需时间明显缩短,细胞数亦明显减少,但囊胚ICM细胞显示AKP阳性,深信其利用价值会得到进一步证实。  相似文献   

11.

Purpose

The objective of this study was to investigate the effects of vitrification on the preimplantation developmental competence of mouse 2-cell, 4-cell and 8-cell stage embryos.

Methods

Mouse 2-cell, 4-cell and 8-cell stage embryos were cryopreserved using the cryotop vitrification method and subsequently warmed on a later date. The embryos were then assessed by their morphology, blastocyst formation and hatching rates. Additionally, trophectoderm (TE) and inner cell mass (ICM) cell numbers were compared in hatched blastocysts from the control and experimental groups.

Results

Vitrified embryos at the 2-cell, 4-cell and 8-cell stages appeared morphologically normal after warming. The overall survival rate of vitrified embryos at various stages after warming was 96.7% and there were no significant differences among 2-cell stage (96.0%), 4-cell stage (96.8%) and 8-cell stage (97.1%) embryos (P?>?0.05). The blastocyst formation rate (69.4%) and hatching rate (52.6%) of vitrified 2-cell embryos were significantly lower than that from the control group and vitrified 8-cell embryos (P?<?0.05). In the vitrified 4-cell embryo group, the blastocyst formation rate (90.3%) was similar to the 8-cell group (91.2%), but the hatching rate (60.0%) was significantly lower than that of the non-vitrified control ( 84.1%) and vitrified 8-cell embryo (78.4%) groups (P?<?0.05). When further development to the fully hatched blastocyst stage was compared, hatched blastocysts derived from vitrified 2-cell, 4-cell and 8-cell embryos had significantly lower cell counts both in the ICM and TE, as compared to fresh blastocysts (P?<?0.05). Among the vitrified 2-cell, 4-cell and 8-cell embryo groups, there were no significant differences in the cell counts of ICM and TE (P?>?0.05).

Conclusions

Although cryotop vitrification was suitable for the cryopreservation of mouse embryos from the 2-cell stage, 4-cell stage and 8-cell stage without significant loss of survival, vitrification had an adverse effect on the development of 2-cell embryos. Mouse embryos at the 8-cell stage had the best tolerance for vitrification and would yield the highest level of post-vitrification developmental competence among early cleavage stage embryos. Nevertheless, it is unclear how these findings can be extrapolated to human embryos.  相似文献   

12.

Purpose

The arrangement of the blastomeres within the 4-cell stage embryo reflects the orientation of the cleavage planes during the second division. To examine their relevance, the developmental capacity and the pregnancy rate were compared between tetrahedral-shaped and non-tetrahedral-shaped 4-cell stage human embryos.

Methods

The study included 3,546 4-cell stage embryos. The arrangement of the blastomeres at the 4-cell stage was annotated as being tetrahedral or non-tetrahedral on day 2 of preimplantation development. Embryo quality was compared on day 3 and day 5. Pregnancy rates were calculated per single embryo transfer on day 3 or day 5.

Results

In total, 2,803 4-cell stage embryos (79 %) displayed a tetrahedral arrangement and 743 (21 %) displayed a non-tetrahedral arrangement. Tetrahedral-shaped embryos developed more into high-quality embryos on day 3 (p < 0.001) and day 5 (p = 0.036) and had a higher blastulation rate (p = 0.009). Though, the number of high-quality embryos selected for transfer did not differ between both groups on day 3 (p = 0.167) and day 5 (p ~ 1). Three hundred thirty single embryo transfers were analysed. No significant difference in clinical pregnancy was found between both groups after transfer on day 3 (p = 0.209) and day 5 (p = 0.653).

Conclusions

The arrangement of the blastomeres according to their previous cleavage planes was correlated to the developmental potential of the 4-cell stage embryo up to the blastocyst stage. If embryo transfers are performed on day 3 and day 5 of development using embryos of adequate quality, the blastomere arrangement at the 4-cell stage had no predictable value regarding pregnancy success.  相似文献   

13.
Objective: To examine the rescue of mouse embryos from 2-cell blocks by the microinjection of maturation promoting factor (MPF) extracted from matured Xenopus eggs into one of the blastomeres of 2-cell stage mouse embryos.

Design: Controlled laboratory study.

Setting: First Department of Obstetrics and Gynecology, Toho University School of Medicine, Tokyo, Japan.

Animal(s): Eight- to 10-week-old female Crj:CD-1(ICR) mice.

Intervention(s): One of the blastomeres of the mouse 2-cell embryos was injected with MPF (MI group) or mHTF medium (MED group) at 28–32 hours after insemination.

Main Outcome Measure(s): The developmental rate to blastocyst.

Result(s): The developmental rate to blastocyst in the MI group (48.0%) was significantly higher than that in the MED group (0%).

Conclusion(s): The 2-cell block was specifically rescued by the microinjection of MPF and not by the insertion of pipettes.  相似文献   


14.
Objective: To determine the most viable embryos for transfer.

Design: Study 1: Preselection of early-cleaving 2-cell embryos for transfer. Study 2: Alternating weeks during which preselection was performed and not performed.

Setting: ART program, Birmingham Women’s Hospital, Birmingham, United Kingdom.

Patient(s): Patients undergoing IVF or ICSI cycles with transfer on day 2.

Intervention(s): Culture of all fertilized embryos.

Main Outcome Measure(s): Number of fertilized embryos cleaving to the 2-cell stage on day 1, embryo quality, implantation rates, and pregnancy rates.

Result(s): Patients with early-cleaving 2-cell embryos had significantly higher pregnancy and implantation rates (45 of 100 [45.0%] and 58 of 219 [25.5%], respectively) than did patients without early-cleaving 2-cell embryos (31 of 130 [23.8%] and 43 of 290 [14.8%], respectively). In weeks during which preselection was used, the overall pregnancy and implantation rates of the clinic improved.

Conclusion(s): The presence of early-cleaving 2-cell embryos improves a patient’s chance of achieving pregnancy. Use of more stringent embryo selection criteria can improve overall pregnancy rates.  相似文献   


15.
The aim of the present study was to examine the effect of culture under 5 and 20% oxygen on the development, differentiation and viability of zygotes and in-vivo-produced embryos at the 2-cell and 8-cell stages of development. First, zygotes collected in a common pool were cultured in 20% O2 for 0, 23, 46 and 95 h. Zygotes and in-vivo-produced embryos at the 2-cell and 8-cell stages of development were then cultured in 5 or 20% O2. The proportion of embryos reaching the compaction and blastocyst stages of development did not differ between groups regardless of the period of time embryos were cultured in 20% O2 or the stage at beginning of culture. Duration of culture under 20% O2 had a significant effect on total number of blastocyst cells. A stage-specific effect was observed on total and trophectoderm cell numbers in blastocysts resulting from the culture of zygotes and in-vivo-produced embryos under 20% O2. ICM and percent ICM development was significantly decreased by culture in 20% O2 at all stages examined. Oxygen concentration had no effect on implantation rate and fetal weights upon embryo transfer. However, transfer of zygotes grown to the blastocyst stage in 20% O2 resulted in a dramatic decrease in fetal development per blastocyst and fetal development per implantation. These results demonstrate that culture of F1 mouse zygotes in 20% O2 compromises the developmental potential of resultant blastocysts, which appear to be normal on morphological assessment.  相似文献   

16.
人卵泡液对小鼠胚胎生长发育影响的研究   总被引:3,自引:0,他引:3  
李娟  张丽红  孟祥阁  张琦  盖凌 《中华妇产科杂志》2001,36(4):215-217,T001
目的:探讨人卵泡液对小鼠胚胎生长发育的影响。方法:在培养液中分别加入人卵泡液(卵泡液组)和人血清(血清组),观察和检测2细胞期鼠胚在体外发育至8细胞期、囊胚期以及鼠胚孵出的比率;用放射免疫方法测定人卵泡液和血清中表皮生长因子(EGF)含量。结果:卵泡液组有72.9%的2细胞期鼠胚能发育到8细胞期,血清组仅有48.0%到达8细胞期;继续培养显示,卵泡液组细胞分裂较快,有50.9%的鼠胚可发育至囊胚期,并有26.3%孵出,明显高于血清组(24.5%,6.9%),两组比较,差异有显著性(P<0.05)。卵泡液中EGF的含量为0.50μg/L,血清中为0.26μg/L,两组比较,差异有显著性(P<0.05)。结论:卵泡液对早期胚胎发育有促进作用,在早期胚胎培养中,添加人卵泡液比添加人血清对提高早期胚胎培养质量更为有效。  相似文献   

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