Effect of an Improved Mechanical Method for Assisted Hatching on the in vitro Development of Mouse Embryos

Objective To evaluate the safety and efficiency of an improved shape of opening for mechanical assisted hatching （AH） on the in virto development of mouse embryos. Methods A total of 622 KM BAI mouse embryos in 2-cell-4-cell stage were randomly divided into group A, group B and control group. A new mechanical AH method by improving the shape of opening in the ZP was used in group A, and a ＂-/ ＂-shaped opening was created. A ＂＋ ＂ -shaped opening was made in group B, while no opening was made in control group. Comparisons have been made among the three groups with regard to the duration of AH, the blastocyst formation and complete hatching rate, etc. Results The duration of AH in group A （43.25 ±3.46 s） was significantly shorter than that in group B （52.81 ±4.32 s, P 〈0.05）. The blastocyst formation rate on d 5 was not significantly different among the three groups （92.27%, 93.66% and 94.92% respectively, P 〉0.05）. The complete hatching rate of blastocysts on d 6 between group A and group B was no statistical difference （94.09% vs 92.71%, P 〉0.05）, but significantly higher than that in control group （43.32%, P 〈0.001）. No significant difference in the percentage of grade 1 blastocysts was found among the three groups on d 5 （85.22%, 82.81% and 86.63% respectively, P 〉0.05）. Conclusion R could enhance the process of embryo hatching and facilitate the hatching rate of blastocysts by using the improved mechanical AH method, which is of safety and efficiency to mouse embryo in the in vitro development.

Effect of royal jelly on in vitro fertilization and early embryo development following nicotine treatment in adult female rats

Objective:To scrutinize the protective role of royal jelly as an antioxidant on nicotine-induced changes in malondialdehyde(MDA)level,p53 expression,in vitro fertilization(IVF)rate,and early embryo development in adult female rats.Methods:A total of 56 adult female Wistar rats were divided into 8 groups(n=7 in each group).Group 1 served as an untreated control group,group 2,3 and 4 received nicotine at a dose of 0.50,1.00 and 2.00 mg/kg respectively,group 5 received royal jelly at a dose of 100.00 mg/kg,and group 6,7 and 8 received 0.50,1.00 and 2.00 mg/kg nicotine,respectively,with 100.00 mg/kg body weight royal jelly.Nicotine and royal jelly were administered daily for 49 days in the experimental groups intra-peritoneally and orally,respectively.At the end of the experimental period,p53 expression,IVF rate and early embryo development as well as MDA concentration were measured.Results:The IVF rate,number of cumulus oocytes,two-cell embryos and blastocysts decreased in the nicotine-treated groups in a dose-dependent manner.In addition,p53 mRNA expression and MDA levels increased in the nicotine-treated groups.Royal jelly co-administration led to partial improvement in the aforementioned parameters.Conclusions:Royal jelly may have a repro-protective effect in nicotine-administered female rats in terms of its anti-oxidant and anti-apoptotic properties.

A Preliminary Observation on the Development of Mouse Embryos Co-cultured with Human Oviductal Tissue or Conditioned Medium in Vitro

The present investigation has been carried out to examine the effect of human oviductaltissue co-culture system on the development of mouse embryos in vitro. Two-cell embryos collected from superovulated mouse were co-cultured with human oviductal tissue suspended inHam 's F10+10% Fetal Calf Serum(F10 FCS),or,in oviductal tissue conditioned medium andF10 FCS as control.The results showed that the proportion developed into blastocyst,proportion of hatchedand the velocity of embryo development were higher in both tissue co-culture and conditionedmedium as compared with F10 FCS control. Furthermore,the velocity and percentage ofembryomic development were higher in co-culture with ampullary tissue or its conditioned medium than that of isthmus.The effects of co-culture and conditioned medium on embryo development had no significant difference. All the embryos obtained from two co-culture systemscould cleave normally.This experimental observation indicated that human oviductalepithelium might secrete some factors to promote the embryonic development in vitro.

Effects of 5-aza-2’-deoxyctidine on the development of porcine parthenogenetic and nuclear transfer embryos

The current study was to investigate whether embryo or fetal fibroblast cells treated with 5-aza-2’-deoxyctidine (5-aza-dC) have a positive effect on the in vitro development of porcine parthenogenetic and cloned embryos. To this end, porcine fetal fibroblast cells were treated with different concentrations (5 nM, 50 nM and 500 nM) of 5-aza-dC for different exposure times (1, 6 and 20 hours), the results showed that DNA methylation in PRE-1 SINE region was gradually reduced over time in cells treated with 5-aza-dC. To determine the effect of 5-aza-dC on in vitro development of porcine activated oocytes, the parthenogenetic embryo was treated with 5-aza- dC. Notably, treatment with 5 nM 5-aza-dC for 1 hour led to a significant improvement in blastocyst development, compared with the control group. The effects of donor cell treatment with 5-aza-dC on porcine cloned embryos development were further examined by treating fetal fibroblast cells with various concentrations (5 nM, 50 nM and 500 nM) of 5-aza-dC for different exposure times (1, 6 and 20 hours). Exposure of cells in 5 nM 5-aza-dC for 1 - 20 hours led to a significant improvement in the percentage of developed blastocysts, while treatment with 500 nM 5-aza-dC did not affect blastocyst development, compared to untreated controls. These findings indicate that treatment of fetal fibro-blast cells with relatively low concentrations of 5-aza-dC for short exposure times improves subsequent blastocyst development of porcine cloned embryos.

Alteration of ERβ gene Rsal polymorphism may contribute to reduced fertilization rate and embryonic developmental competence

This paper aims to determine the possible role of estrogen receptor-β （ERβ） gene Rsal polymorphism on sperm fertility and early embryonic development in humans. Three groups of Chinese men were recruited： in vitro fertilization （IVF） group, including 374 couples who underwent conventional IVF; intracytoplasmic sperm injection （ICSI） group, including 294 couples who underwent an ICSI procedure using ejaculated sperm; and azoospermic group, consisting of 197 couples who underwent ICSI using either testis or epididymis sperm. Rsal polymorphism in the ERβ gene was detected by polymerase chain reaction （PCR）-restriction fragment length polymorphism technique; fertilization and high-quality embryo rates were evaluated for each group. In each group, no significant differences were found in the overall rates of fertilization and high-quality embryos among GG, AG and AA genotypes. However, the proportion of cycles possessing a satisfactory high-quality embryo rate with the AA genotype was significantly lower than that in the wild-type GG genotype from each group. These results demonstrated that sperm possessing the ERβ RsalA genotype may have reduced fertilization ability and decreased early embryonic developmental potential, which could directly or indirectly contribute to the low fertilization rate and early embryonic developmental arrest in some cases.

Effect of Insulin on Development of ICR Mouse Embryos in Vitro

Objective To study the effects and the appropriate concentration of insulin on the development of mouse embryos in vitro, and the effects of insulin on the development of different stages of embryos.Methods Mouse embryos were cultured in vitro in the mKSOM media supplemented with insulin at different concentrations and with insulin during different stages of embryos. The blastoeyst rates and the cell numbers were counted.Results Additions of insulin significantly increased the rates of blastocyst and the total cell numbers. The concentrations of 0.005 μg/ml and 0.05 μg/ml insulin caused a significant increase in the total cell numbers compared with the control and experimental groups. Addition of insulin from 2-cell to 4-cell stage or from the 4-cell to morula stage, significantly increased the blastocyst rates compared with control and experi- mental groups.Conclusion Insulin can promote the development of mouse embryos in vitro. The appropriate concentration of insulin added in mKSOM was 0.005 μg/ml and 0.05μg/ml. Exposure of embryos to insulin, beginning at the 2-cell and extending to the 4-cell stage or beginning at the 4-cell and extending to the morula stage, is important for the development of lCR mouse embryos in vitro.

In-vitro Developmental Potential of Single Blastomeres Derived from Day 3 Discarded Human Embryos

Objective To investigate whether isolated blastomeres of discarded human embryos could develop into blastocysts cultured in vitro without zona pellucida. Methods Total discarded 60 embryos, which were not suitable for transplanting or freezing, were collected from 21 patients. Of 60 embryos, 10, 8, 24, 12, and 4 embryos were at 2-, 3-, 4-, 5- and 6-cell stage, respectively. These embryos were split by 0.5% protease combined with mechanical method. The resulting single blastomere was cultured individually, evaluated daily and counted blastocyst development. Pluripotency of inner cell mass （ICM） of the blastocyst was analyzed by the expression of alkaline phosphatase（AKP）. Results A total of 229 single blastomeres were isolated from 60 embryos. Defining the day when the embryos were split as the first day （d 1）. The majority of the blastomere- derived embryos followed the normal pattern of development with compaction on d 3 and cavitationon d 4 and developed into small blastocysts on d 5. Rates of division, compaction, cavity and expansion of single blastomeres from 4-cell embryos were higher than those in other groups （P 〈O.05）, and AKP was expressed in ICM. Conclusion Some of the blastomeres from discarded human embryo are flexible and able to develop into blastocysts, the potential was related to their donor embryos closely. They seem to follow development pattern of their donor embryos. The blastomere- derived blastocysts have smaller size and fewer cells compared with regular in vitro cultured human embryos. However, AKP expression showed ICM cells with pluripotency. We believe that the value of single blastomere of discarded embryos will be further confirmed in the future.

不同培养基对小鼠卵母细胞体外成熟质量及发育潜能的影响

背景:近年,未成熟卵母细胞体外成熟技术的需求逐渐增加。卵母细胞成熟受多种因素影响,其中培养基的选择尤为重要,目前尚无统一方案。目的:用不同成熟培养基对生发泡期卵母细胞体外成熟,研究其对卵母细胞质量及发育潜能的影响。方法:用G-1^(TM)PLUS培养基、CZB培养基和M16培养基对生发泡期卵母细胞体外成熟,并以体内成熟卵母细胞为对照组,比较各组间的体外受精和早期胚胎发育能力。对于各组成熟后卵母细胞,采用免疫荧光方法评估线粒体功能,共聚焦显微镜实时成像系统检测钙震荡情况。结果与结论:①3组之间第一极体排出率无明显差异(P>0.05);②G-1^(TM)PLUS组体外受精率(52.86±11.24)%高于M16组(37.76±6.70)%和CZB组(30.62±5.51)%;CZB组的囊胚率(36.23±6.63)%低于对照组(78.16±4.17)%、G-1^(TM)PLUS组(55.75±7.63)%和M16组(53.36±6.33)%;③与对照组相比,仅CZB组的纺锤体长宽比增加;④CZB组线粒体功能差于对照组、G-1^(TM)PLUS组及M16组,并出现CZB组线粒体异常凝集现象;⑤受精后CZB组及M16组的钙震动频率显著高于G1组和对照组。综上所述,在小鼠卵母细胞体外成熟过程中,G-1^(TM)PLUS、CZB及M16培养基的体外成熟率无差异,但G-1^(TM)PLUS培养基有更高的受精率和囊胚形成率。

抗着丝点抗体阳性患者体外受精结局分析

目的:通过观察抗着丝点抗体(ACA)阳性患者体外受精-胚胎移植和自然试孕结局,探讨此类患者的生育策略。方法:采用病例对照研究回顾性分析2016年6月至2023年6月在浙江省人民医院接受体外受精-胚胎移植治疗且有抗核抗体(ANA)谱检查结果的3955例患者的临床资料。根据ACA结果将所纳入患者分为ACA阳性组和ACA阴性组。采用倾向评分匹配方法对两组进行1∶3配对,分别比较两组体外受精的胚胎结局;并采用自身对照分析不同授精方法和是否应用免疫抑制剂对结局的影响;对ACA阳性患者体外受精失败后的自然试孕和疾病进展进行随访。结果:ACA阳性患者34例,占总病例数的0.86%,占ANA阳性体外受精患者数的2.51%。无论是接受常规体外受精(c-IVF)还是卵胞质内单精子注射(ICSI)的患者,ACA阳性组卵母细胞成熟度和受精情况均与ACA阴性组有明显差异(均P<0.01),且ACA阳性组授精后第三日(D3)次优胚数和D3优胚数均减少(均P<0.05)。5例ACA阳性患者自身ICSI周期相比c-IVF周期双原核(2PN)率未提高(P>0.05),D3优胚数和D3次优胚数减少(均P<0.05)。12例ACA阳性患者经过免疫抑制剂治疗1~2个月后再行c-IVF/ICSI,用药前后获卵和受精情况均未改变(均P>0.05),2PN胚胎卵裂率改善(P<0.05)。ACA阳性组与ACA阴性组移植胚胎数相近,但ACA阳性组胚胎着床率、临床妊娠率显著低于ACA阴性组(均P<0.05),流产率差异无统计学意义(P>0.05)。27例ACA阳性患者体外受精失败后尝试自然试孕或人工授精,共获临床妊娠7例。结论:血清ACA阳性会干扰卵母细胞的成熟和正常受精过程,使用ICSI和免疫抑制剂不能改善受精结局,但ACA阳性患者有可能获得自然妊娠。

精子DNA碎片对体外受精-胚胎移植妊娠结局及胚胎发育的影响

目的:分析精子DNA碎片指数(DFI)对体外受精-新鲜胚胎移植(IVF-ET)助孕及活产结局的影响。方法:选取2015年6月至2020年12月因女方因素首次行IVF并进行新鲜胚胎移植的患者共947例,根据DFI分为低、中、高3组:DFI<15%、15≤DFI≤30%和30%<DFI,比较3组双方基础数据,女方包括年龄、体质量指数(BMI)、基础生殖激素、AMH等;男方包括年龄、BMI、精液参数;超促排卵参数;处理前和处理后IVF精液参数;促排卵结局、胚胎质量及妊娠结局。结果:不同DFI水平间相比,双方年龄在3组间均有显著性差异,15≤DFI≤30%和30%<DFI组双方年龄显著大于DFI<15%组;基础状态精液参数中前向运动精子百分率、非前向运动精子百分率、不活动精子百分率,以及IVF处理前精子总活率、前向运动精子百分率在3组间也均有显著差异(P<0.01);中和高DFI组中基础精子正常形态率、IVF处理前精子浓度、精子总数和处理后精子浓度均显著低于低DFI组;30%<DFI组无论是精液处理前还是精液处理后精子总数均显著低于其他两组(P<0.01)。此外,中和高DFI组受精率显著低于低DFI组,其他临床数据包括促排卵参数、胚胎质量、妊娠结局组间均无显著差异。结论:高DFI水平对精子活率产生显著影响,IVF精液处理后DFI水平与精子浓度、精子总数、受精率均呈显著负相关,对胚胎质量及妊娠结局无影响。

体外成熟卵母细胞质量和胚胎发育潜能影响因素的研究进展

随着生殖医学的迅速发展,卵母细胞体外成熟(IVM)技术取得了显著进步,但目前IVM卵母细胞仍然面临成熟率低、受精率低和受精后胚胎发育潜力低等挑战。本文总结影响IVM卵母细胞质量、受精及胚胎发育潜能的因素,旨在为增强卵母细胞IVM能力和胚胎发育潜能提供创新思路。

男性性激素与抗米勒管激素水平对体外受精-胚胎移植胚胎发育的影响

目的探讨男性性激素与抗米勒管激素(AMH)水平对体外受精-胚胎移植(IVF-ET)胚胎发育的影响。方法回顾性分析2015年5月至2021年8月于西北妇女儿童医院接受体外受精(IVF)/卵胞质内单精子注射(ICSI)助孕的189例男性患者的临床资料,根据受精卵受精72h后是否有优质胚胎将其分为优质胚胎组和无优胚胎组。对比两组临床资料,通过LASSO和Logistic回归模型探讨影响优质胚胎形成的相关因素,采用受试者工作特征(ROC)曲线评价及性激素水平对胚胎发育情况的预测价值,构建列线图模型并进行评价。结果无优胚胎组的睾酮(T)、雌二醇(E_(2))、AMH、精子密度、精子活动率、精子速度试验结果、生育力指数、活动力指数低于优质胚胎组(P<0.05)。多因素分析结果显示,E_(2)、T、AMH水平及精子速度试验结果、生育力指数均为患者胚胎发育的独立影响因素(P<0.05)。与E_(2)、T及AMH单独预测相比,三者联合的预测效能明显较高。构建的列线图模型具有较高的区分度、准确性和临床适用性。结论血清E_(2)、T、AMH水平异常变化与男性不育症患者无优胚胎形成密切相关,三者联合对IVF-ET胚胎发育具有较高的预测价值,可作为临床筛选高危人群的敏感指标。

重组卵泡刺激素预充注射笔在控制性超促排卵中的临床应用

目的探讨重组卵泡刺激素预充注射笔在中国人控制性超促排卵中的效果与可行性。方法对南方医科大学南方医院生殖医学中心2007.01～2008.04间就诊的184例需行体外受精-胚胎移植(IVF-ET)不孕症妇女,采用黄体期长方案,随机分别给予重组卵泡刺激素预充注射笔(A组)和重组卵泡刺激素冻干粉制剂(B组)进行卵巢刺激,分析比较两组间用药情况、促排卵与IVF-ET结局。结果A组rFSH总用量、前往医疗单位次数、购药和注射费用明显少于B组(P<0.05);在用药时间上没有显著性差异(P>0.05);A组剩余药量明显高于B组(P>0.05)。A组用药局部红肿的比例显著少于B组(P<0.05),但用药局部瘀斑的比例明显多于B组(P<0.05)。A组药物易用性和方便性评分均显著高于B组(P<0.05);两组在疼痛感受上没有显著性差异(P>0.05)。A组进行了更频繁的药物个体化调整(P<0.05),而其hCG日血清E2浓度明显低于B组(P<0.05)。但两组hCG日内膜厚度、总获卵数、第二次减数分裂中期卵子数量、每周期移植胚胎数以及临床妊娠率没有显著性差异(P>0.05)。A组成熟卵子比例、优质胚胎率和胚胎着床率明显高于B组(P<0.05),而发生中重度OHSS的例数明显少于后者(P<0.05)。结论重组卵泡刺激素预充注射笔具有高效、安全、易用、价廉和患者耐受性好等优点,有利于个体化COS治疗方案的实施,在临床推广中应注意合理选择和搭配制剂种类并加强对患者的健康教育。

HA对牛卵母细胞体外成熟和早期胚胎体外发育影响的研究

在TCM-199中添加一些确定的辅助成分,探讨在无血清培养基中添加不同浓度HA对牛卵母细胞体外成熟及早期胚胎体外发育的影响。结果表明,在卵母细胞成熟培养液(TCM-199-m)中添加4.0mg/mL的HA时,卵母细胞的成熟率和卵裂率与对照组(BSA)无显著差异(P>0.05),分别为85.14%,83.57%和89.47%,85.24%,但显著高于其他各浓度组(P<0.05),表明HA在卵母细胞无血清成熟培养中可代替BSA。在受精卵体外培养液(TCM-199-c)中添加HA时,囊胚发育率与对照组(BSA)差异不显著(P>0.05),分别为27.64%和26.51%,但显著高于TCM-199-c组(P<0.05),表明HA对牛胚胎体外发育具有明显的促进作用,在无血清培养中可以代替BSA。当在TCM-199-c+HA中添加少量的BSA时,囊胚发育率(31.19%)显著高于其他各组(P<0.05),表明HA欲完全代替BSA还有不完善之处,有待进一步研究。

卵泡发育速度对胚胎发育及体外受精-胚胎移植结局的影响

目的探讨促排卵过程中卵泡发育速度与卵母细胞质量间的相关性。方法154名患者共159个体外受精-胚胎移植(IVF-ET)周期,按卵泡发育速度(v,mm/d)的分布分为A组(v<1)、B组(1≤v≤1.59)、C组(v>1.59),比较各组卵母细胞成熟度指标。结果A、B、C3组卵细胞成熟率分别为89.56%、92.45%和94.76%,优质胚胎率分别为40.85%、36.37%和37.11%,妊娠率分别为42.1%、43.1%和35.3%,卵母细胞成熟率C组高于A、B组,优质胚胎率A组高于B、C两组,但组间差异均无显著性(P>0.05),而妊娠率A、B两组明显高于C组,且B、C两组间差异有显著性(P<0.05)。结论促排卵过程中卵泡的生长速度应控制在一个最佳的范围内,因其与卵子质量和后期优质胚胎率及妊娠结局密切相关,且卵泡发育速度又可作为衡量卵泡发育潜能的一项重要指标。

绵羊体外成熟、体外受精卵的体外发育及移植后的产羔

体外成熟、体外受精后的绵羊卵在体外长期培养或在2～4细胞期和桑堪～囊胚期移植给受体母羊,观察了培养卵的发育和移植后的受胎率。方法是将采自屠宰场的绵羊卵巢在1～12小时内带回实验室,抽取卵母细胞。从中选取卵丘细胞层完整的卵子,在二氧化碳培养箱内,用含有10％NSS(或FCS)、hCG和E_2,并以Hepes缓冲的M199培养24～26小时。再用以IonophoreA23187诱导获能处理过的新鲜公羊精于进行体外受精.7～10小时后移入发育培养基,即含有10％FCS(或NSS)和丙酮酸钠的Hepes缓冲的M199内继续培养,受精后将部分发育为2～4细胞胚和桑堪～囊胚期胚手术移植给受体母羊。另一部分卵子则在受精后48～72小时的不同时间内统计其卵裂卵的出现率,并继续培养7～12天详细观察卵裂卵的发育情况。在受精后48～72小时卵裂卵的出现率,FCS和NSS组分别为39.6％145/366)和52.4％(182/347),前者显著低于后者;将61枚2～4细胞期胚和桑椹～囊胚期胚分别移植给20只受体母羊,有10只受胎。共产羔11只,受胎率为50％;在发育培养液内继续培养的482枚卵裂卵中有312枚(64.7％)发育为桑椹～囊胚期胚(其中包括部分孵化囊胚)。

葡萄糖对ICR小鼠胚胎体外发育的影响

研究葡萄糖在小鼠早期胚胎体外发育中的作用。实验1将6—8周龄的ICR雌鼠超数排卵后与公鼠交配,收集1-细胞放入含0(对照组)、0.5、1、3、5、10mmol/L葡萄糖的CZB中培养;实验2将从超排的ICR雌鼠输卵管内收集的1-细胞放入无糖CZB中培养,分别于1细胞、2细胞、4细胞、桑椹胚阶段移入含3.0mmol/L葡萄糖(最适浓度)的CZB中,培养24h后又移回到无糖CZB中(桑椹胚阶段除外)继续培养以及整个胚胎培养过程均在含糖CZB中,对照组胚胎培养全程均在无糖CZB中。每组胚胎于37℃、5%CO2培养箱中培养120h,每24h在倒置显微镜下观察胚胎发育情况,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。结果显示,小鼠胚胎在含糖CZB中与在无糖CZB中4-细胞发育率无差异;含糖CZB中囊胚率显著高于对照组;3.0mmol/L浓度组囊胚细胞数显著高于其余组;2-细胞至4-细胞、4-细胞至桑椹胚前添加葡萄糖囊胚率显著高于对照组,1-细胞至2-细胞、桑椹胚及其以后阶段添加葡萄糖囊胚率与对照组无差异。实验证实,在ICR小鼠胚胎体外培养中加入葡萄糖不会导致2-细胞阻滞;葡萄糖浓度增至10mmol/L对ICR小鼠胚胎无毒性作用;ICR小鼠胚胎体外培养的最适葡萄糖浓度为3.0mmol/L;2-细胞至4-细胞、4-细胞至桑椹胚前添加葡萄糖是必要的。

波纹龙虾胚胎的离体培养及发育观察

对波纹龙虾不同发育时期的胚胎进行离体培养和显微观察。结果表明,初始培育的离体胚胎愈接近孵化期,其孵化率愈高(最高达80.0%);波纹龙虾胚胎发育经历受精卵、卵裂期、囊胚期、原肠期、中眼色素形成期、复眼色素形成期、心跳期、破膜前期和出膜期;胚胎在发育过程中颜色变化:橙红→深橙红→砖红→灰白→接近透明。

青钱柳离体胚的培养及快速繁殖

以青钱柳离体胚为材料,采用正交试验设计对影响其萌发、快速成苗的各种因素进行研究,并进行了诱导增殖试验。结果表明,基本培养基种类是影响青钱柳离体胚萌发和成苗最关键的因素。离体胚萌发的最佳培养基组合为WPM+GA3(10 mg/L)+蔗糖(30 g/L),萌发率可达100%;适宜成苗的培养基组合是WPM+GA3(5 mg/L)+麦芽糖(30 g/L),培养60 d的无菌苗移栽成活率为100%;最适的增殖培养基为WPM+6BA(0.5 mg/L),平均增殖系数为3.45。

不同品系小鼠胚胎玻璃化冷冻保存的比较研究

目的 研究甘油作为冷冻保护剂、不同基因型小鼠对胚胎玻璃化冷冻的影响。方法 采用 6 5mol L的甘油作为冷冻保护剂 ,采用二步法对CBA、NOD、C57BL 6J、ICR及CD1小鼠 3 5d的胚胎进行玻璃化冷冻 ,并比较了不同品系小鼠胚胎的复苏率及移植受孕率。结果和结论 CBA、NOD、C57BL 6J,ICR及CD1的复苏率分别为 5 7 6 %、4 8%、31 3%、86 5 %及 88% ,移植受孕率为 2 1%、2 3 5 %、11%、38%和 35 5 % ,封闭群小鼠的胚胎复苏率、移植受孕率均显著高于近交系小鼠。这提示胚胎的复苏率及移植受孕率可能与小鼠的不同基因型有关。五个品系中 ,桑椹胚及早期囊胚的体外复苏率均显著高于扩张囊胚。