A New Micropropagation Technology of Tilia amurensis:In VitroMicropropagation of Mature Zygotic Embryos and the Establishment of a PlantRegeneration System

Tilia amurensis is an economically valuable broadleaf tree species in Northeast China.The production of highqualityT.amurensis varieties at commercial scales has been greatly limited by the low germination rates.Thereis thus a pressing need to develop an organogenesis protocol for in vitro propagation of T.amurensis to alleviate ashortage of high-quality T.amurensis seedlings.Here,we established a rapid in vitro propagation system forT.amurensis from mature zygotic embryos and analyzed the effects of plant growth regulators and culture mediain different stages.We found that Woody plant medium(WPM)was the optimal primary culture medium formature zygotic embryos.The highest callus induction percentage(68.76%)and number of axillary buds induced(3.2)were obtained in WPM+0.89μmol/L 6-benzyladenine(6-BA)+0.46μmol/L kinetin(KT)+0.25μmol/Lindole-3-butryic acid(IBA)+1.44μmol/L gibberellin A_(3)(GA_(3)).The multiple shoot bud development achievedthe highest percentage(83.32%)in the Murashige and Skoog(MS)+2.22μmol/L 6-BA+0.25μmol/L IBA+1.44μmol/L GA_(3).The rooting percentage(96.70%)was highest in 1/2 MS medium+1.48μmol/L IBA.Thesurvival percentage of transplanting plantlets was 82.22%in soil:vermiculite:perlite(5:3:1).Our study is the firstto establish an effective organogenesis protocol for T.amurensis using mature zygotic embryos.

Callus Induction from Mature Embryos of Maize

In this study we studied the factors influencing the callus induction from mature embryos of maize inbred lines Qi 319, Zhen 58, Chang 7 -2, Lx 9801 and 81162, such as genotype, combination of plant growth regulators, and low-temperature pretreatment. The results showed that the induction rate of Qi 319 was the highest among the four genotypes tested; combination of 4.0 mg/L 2,4-D ＋ 0.5 mg/L 6-BA was suitable for inducing callus from mature embryos; three days of 4℃ pretreatment can promote the callus induction significantly. The indices optimized in the present study are helpful for establishing genetic transformation system in maize without considering seasonal variation.

Effects of Genotypes and Basic Medium on Culture of Maize Mature Embryos

[Objective]This study was to screen out suitable genotypes and basic medium for the culture of maize mature embryos.[Method]Using mature embryos of nine maize genotypes as explants,the effects of genotypes and basic medium on callus induction and subculture were investigated.[Result]The genotypes performed better in callus induction and subculture were found in turn 853-35,853-209,Dan 34 and 81162.MS medium is better than N6 medium in the callus induction from maize embryos,while N6 medium is more suitable for callus subculture.[Conclusion]Our study further improved the tissue culture system in maize with mature embryos as explants.

Establishment of a Highly Efficient Regeneration System for the Mature Embryo Culture of Wheat

Establishment of a highly efficient regeneration system for the mature embryo of wheat will provide a convenient tool for wheat tissue culture and transformation, thereby facilitating the transformation of foreign genes into wheat. By using the mature embryos derived from 20 different wheat lines including Shi 4185, Yumai 66, Lunxuan 987, CB037, Yangmai 6, Xinchun 9, Bobwhite, Han 6172, Zheng 9023, Jimai 20, Ningchun 4, and Jing 411, the effects of some factors including inoculation methods, initiating culture media, organic additives, antioxidants, and auxins on the regeneration from the explants were evaluated. The results indicated that the scraping embryo culture was better than the whole embryo culture, the Aa medium was better than the SD2 medium and dicamba was better than 2,4-D in increasing the regeneration frequency. An Adi medium was established in this study by adding silver nitrate, cysteine, ascorbic acid, dicamba, glutamine into the Aa medium at the concentration of 4,40, 100, 2, and 5 mg L^-1, respectively. By using the Adi medium and the scraping technique, the regeneration frequencies of the mature embryos of CB037, Lunxuan 987, Hart 6172, Yangmai 6, Bobwhite, Zheng 9023, Shi 4 185, and Jimai 20 became 85.6, 60,1, 46.0, 42.1,42.0, 34.0, 33.0, and 32.0%, respectively, which were about 5-8 times higher than that obtained from the conventional culture mediums and techniques. This novel regeneration system could be helpful in wheat transformation.

Study on Plant Regeneration of Wheat Mature Embryos Under Endosperm-Supported Culture

To reveal the suitability of using mature embryos as an explant source in wheat tissue culture, mature embryos from eight common wheat cultivars （Triticum aestivum L. cv.） were cultured with or without endosperm to test their efficiency of callus induction and plant regeneration. When embryos were cultured together with endosperm （endosperm-supported culture, ES）, the percentage of callus induction was significantly lower than that when embryos were cultured in the absence of endosperm （non-endosperm-supported culture, NES）. This pattern was evident in most genotypes, regardless of whether 2 or 8 mg L^-1 2,4-D was added in the NES culture. However, in ES culture, more induced calli were differentiated into distinct green spots and they further developed into plantlets. Thus, more plants were regenerated in ES culture than in the NES treatment. Most of the eight tested genotypes showed a significant difference in callus induction rate and plantlet regeneration in both ES and NES cultures. In addition, the enzymatic activity of oxalate oxidase in the callus of ES culture condition was obviously higher than that in the callus of NES culture condition, suggesting that the activity of oxalate oxidase may be a parameter for selection of calli with potential for plantlet regeneration. These results indicate that wheat mature embryos are valuable explants for highly efficient callus induction and plant regeneration, if proper treatment and medium are used.

Dicamba and Sugar Effects on Callus Induction and Plant Regeneration from Mature Embryo Culture of Wheat

To establish a highly efficient plant regeneration system for wheat genetic transformation, the effects of three different concentrations of dicamba and two different sugar types on callus induction and plant regeneration from mature embryo cultures were evaluated. Callus induction and plant regeneration were obtained from mature embryos of two commercial cultivars Zhoumai 18 and Yumai 34 （Triticum aestivum L.） cultured on L3 basal medium. The results showed that the efficiency of mature embryo culture was significantly influenced by the genotypes, sugar types and dicamba concentrations. 4 mg L^-1 dicamba proved the best effective for inducing embryogenic callus and also gave the highest proportion of plants regenerated across the two cultivars. Substitution of maltose by sucrose significantly improved the plant regeneration efficiency in both cultivars. There was a significant interaction between genotype-by-sugar types, and sugar types-bydicamba concentrations. Overall, Zhoumai 18 gave the highest frequency of plant regeneration （82.65%） when dicamba concentration was 4.0 mg L^-1 and with sucrose in initial callus induction. These results will facilitate genetic transformation work with elite wheat.

The Comparison in Tissue Culture Ability of Mature Embryo in Different Cultivars of Rice

In order to study the regeneration technology of mature embryos in different rice varieties,nine japonica,nine indica and eleven hybrid rice varieties of two line or three line or superiority combinations were selected as explants to study the callus induction,differentiation and regeneration rates on different media.The higher callus induction （61.7-89.2%） was observed in japonica rice,when cytokinin was added at lower concentration （0.3 mg L-1 6-BA） in M8 basal medium,supplemented with 30 g L-1 sucrose,8 g L-1 agar and 2 mg L-1 2,4-D.Further,the addition of two cytokinins （2 mg L-1 6-BA,0.5 mg L-1 KT） and 1 mg L-1 NAA in the M8 basal supplemented medium resulted in 9.1-100% of the callus induction in indica rice.The percent callus induction in hybrid rice varieties was 40-86.3% when addition of 1 mg L-1 6-BA and 1 mg L-1 KT was added,and the cytokinins was required by the japonica and indica rice varieties in the M8 basal supplemented medium.It was observed that when the 0.5 mg L-1 2,4-D and 1 mg L-1 6-BA were added in japonica rice,and 0.2 mg L-1 2,4-D and 0.5 mg L-1 6-BA were added in indica and hybrid rice in the MS different media,the regeneration rates were 9.2-59.5%,3.6-87.5% and 17.2-43.2% for japonica,indica and hybrid rice,respectively.Thus,the regeneration technology with higher output is established in the mature embryos of similar rice varieties.

Establishment and Optimization of the Regeneration System of Mature Embryos of Maize (Zea mays L.)

A reliable system was developed for regeneration from mature embryos derived from callus of four maize inbred lines （Liao 7980, Dan 9818, Dan 340, and Dan 5026）. The protocol was mainly based on a series of experiments involving the composition of culture medium. We found that 9 pM 2,4-dichlorophenoxyacetic acid in MS medium was optimum for the induction of callus. The induction frequency of primary calli was over 85% for four inbred lines tested. The addition of L- proline （12 mM） in subculture medium significantly promoted the formation of embryogenic callus but it did not significantly enhance growth rate of callus. Efficient shoot regeneration was obtained on regeneration medium containing 2.22 μM 6- benzylaminopurine in combinations with 4.64 μM Kinetin. Regenerated shoots were rooted on half-strength MS medium containing 2.85 μM indole-3-butyric acid. This plant regeneration system provides a foundation for genetic transformation of maize.

Transcript Profiling Reveals Abscisic Acid,Salicylic Acid and Jasmonic-Isoleucine Pathways Involved in High Regenerative Capacities of Immature Embryos Compared with Mature Seeds in japonica Rice

Induced pluripotent cell mass plays a role in genetic transformation mediated by Agrobacterium. Mature seeds are more recalcitrant to the induction of suitable calli than immature embryos in rice, but the exact molecular mechanisms involved remain elusive. In this study, the morphological structure of calli induced from mature seeds and immature embryos were observed under a scanning electron microscope using a paraffin embedded technique. Meanwhile, a total of 2 173 up- and down-regulated genes were identified in calli induced from mature seeds and immature embryos by RNA-seq technique and furtherly confirmed by quantitative real-time PCR. The results revealed the remarkable morphological differences in calli induced from mature seeds and immature embryos, and plant hormone signal transduction and hormone biosynthesis pathways, such as abscisic acid, salicylic acid and jasmonic-isoleucine, were found to play roles in somatic embryogenesis. This study provided comprehensive gene expression sets for mature seeds and immature embryos that were served as an important platform resource for further functional studies in plant embryogenesis.

Effects of CuSO_4 and Uniconazole on Mature Embryo Culture in Japonica Rice

In order to establish the system of high frequency plant regeneration for japonica rice mature embryos, the effects of different concentrations of CuSO4 and uniconazole on in vitro culture of mature embryos were studied using three rice cultivars of Kongyu 131, Longjing 24, and Dongnong 425 as test materials. The results showed that callus induction and differentiation of japonica rice mature embryos were apparently improved on the medium with 10-15 μmol·L-1 CuSO4 and 0.50-1.00 mg·L-1 uniconazole. Induction and differentiation rates of different genotype rice mature embryos displayed different sensitivities to CuSO4 and uniconazole. For the callus induction frequency of three varieties, the optimal concentration of CuSO4 was 15.0 mol·L-1. When the concentration of CuSO4 was 15 μmol·L-1, the plantlet differentiation rates of Kongyu 131 and Dongnong 425 got to the highest, while the concentration of CuSO4 was 10 μmol·L-1 for Longjing 24. For the callus induction and plantlet differentiation rates of Kongyu 131 and Dongnong 425, the ideal concentration of uniconazole was 0.50 mg·L-1 and for Longjing 24 was 1.00 mg·L-1.

Plantlet regeneration from mature zygotic embryos andembryonic explants of masson pine (Pinus massoniana Lamb.)

Excised zygotic embryos, cotyledons and hypocotyls of juvenile seedlings of masson pine were grown on DCR medium supplemented with several concentrations of various plant phytohormones. BA (1.0 mg/ L) in combination with NAA (0.05 mg/L)in DCR medium was found to increase the formation of adventitious buds from mature zygotic embryos, but most of them were formed at the tips of embryonic cotyledons. Adventitious buds were obtained from cotyledons and hypocotyls from juvenile seedlings when they were cultured on DCR medium containing BA 3-5 mg/L and NAA 0.1-0.2 mg/L. Elongation of buds were observed on hormone-free DCR medium with or without activated charcoal (0.5%). Root initiation was achieved with full or half strength DCR inedium supplemented with IBA 1.0 mg/L and NAA 0.25-0.5 mg/L. Approximately 11-20 axillary buds formed on each explant when juvenile seedling explants were treated (3-20h) with BA 50-100 mg/L, followed by transfer to hormone-free DCR medium. The maximum number of shoots obtained per explant within six months was 33.

Screening and verification of the factors influencing somatic embryo maturation of Larix olgensis

With embryogenic callus of Larix olgensisis, we investigated the effects of inositol, glutamine, casein hydrolysate, carbohydrate, abscisic acid and silver nitrate concentration on the maturation of the somatic embryo.Three dominant factors emerged, and we developed a response surface model based on the Box-Behnken design.We defined the optimal conditions for the maturation of somatic embryos. The contents of abscisic acid, silver nitrate, sucrose and casein hydrolysis significantly affected the amount of maturing embryos, but inositol, maltose and glutamine had no effect. By establishing a response surface model with multiple factors, we predicted that the optimal number of L. olgensis somatic embryos was 204 ± 4 gon basal medium, containing 18.28 mg Labscisic acid,5.46 mg Lsilver nitrate and 82.67 g Lsucrose. In the verification experiments, the addition of 20 mg Labscisic acid, 5 mg Lsilver nitrate and 80 g Lsucrose to BM yielded an average of 202.06 somatic embryos per gram. These results should guide large-scale breeding of L. olgensis.

Effect of L-carnitine supplementation during in vitro maturation and in vitro culture on oocyte quality and embryonic development rate of bovines

Objective:To assess the effect of L-carnitine supplementation during in vitro oocyte maturation and in vitro culture process of bovine oocytes.Methods:L-carnitine(3.8 mM)was added to maturation medium and the effect was assessed in the quality(Experiment 1)and in the cleavage and 4-cells stage(Experiment 2).Besides,the effect of L-carnitine addition on maturation medium(3.8 mM)and culture medium(1.5 mM)on embryo rate production was assessed.In Experiment 1,bovine oocytes from abattoir were randomly separated into two groups(the control group and L-carnitine group)forin vitro maturation.Matured oocytes were examined for cumulus cells expansion as an indicator of maturation,and the content of the mitochondrial activity,the presence of lipid droplets,the reduced glutathione,and the reactive oxygen species were measured by using specific fluorochromes.In Experiment 2,oocytes were matured as performed in Experiment 1,afterward fertilized and cultured until day 3,and cleavage rate and 4-cells stage rate were determinated.In Experiment 3,in vitro maturation and fertilization were done as performed in Experiment 2,but at day 3 of culture,each group of embryos was separated into two new groups,and L-carnitine(1.5 mM)was added in culture media until day 8.The cleavage and embryo development rate were determined on the basis with the oocytes put on maturation.Hatching rate was calculated from cleaved embryos.Results:The cumulus expansion rate at gradeⅢand mitochondrial activity were significantly higher in the L-carnitine group in comparison with the control group(P0.05).In addition,cleavage and the proportion of embryo development and hatching rate were similar for all groups(P>0.05).Conclusions:L-carnitine as a supplement in culture media improves the cumulus expansion and increases the mitochondrial activity during in vitro maturation process but has no apparent effect on the cleavage and development of bovine embryos.Further investigations of L-carnitine addition on in vitroculture are needed to test their effect on embryo quality.

Effect of GPAG Supplement on Porcine Oocyte Maturation and Its Following Embryonic Development by Parthenogenetic Activated

The aim of this study was to compare the effect of GPAG and commonly used FCS on porcine oocyte maturation and subsequent embryonic development after the fertilization. COCs were aspirated from follicles and cultured for 16, 24, 32, 40 and 48 h in TCM-199 medium either with GPAG or FCS. After 24 h with GPAG, 89.4% of oocytes reached M Ⅰ stage while in the medium supplemented with FCS, only 27.7% of oocytes reached the same stage （P〈0.05）. Prolonged incubation for up to 32 h clearly demonstrated that some of oocytes cultured in GPAG medium were at M Ⅱ stage （35.7%）, few of oocytes from FCS medium were at M Ⅱ stage （7.5%） （P〈0.05）. Both groups of oocytes reached the same stage of maturation within 48 h. After 48 h of culture, the oocytes with extruded polar bodies were inseminated. Fertilized oocytes were cultured in PZM3 medium supplemented with 3 mg.mL of BSA. After 7 days, the development and the quality of embryos were evaluated. The results showed that the maturation of oocytes in the presence of GPAG significantly increased their subsequent developmental ability when compared with FCS supplementation （29.2% ： 18.9% of blastocysts, P〈0.05）. However, differential staining revealed that once blastocysts were formed in either group, they had the same total cell number （39 ： 38） and the ICM/total cell ratio （0.26 ： 0.28）

银杉愈伤组织诱导植株再生的影响因素

[目的]探索银杉成熟胚愈伤组织诱导植株再生的影响因素,为阐明影响诱导银杉愈伤组织、不定芽等的机制提供帮助。[方法]以广西金秀和广西桂林2个种源的银杉成熟胚为试验材料,利用1/2 MS、DCR、P6培养基,分别添加0.5、1.0、1.5 mg·L^(−1)的6-BA,0.2、0.4、0.6 mg·L^(−1)的NAA和0.5、1.0、1.5 mg·L^(−1)的IBA,诱导银杉愈伤组织、不定芽和不定根,得到银杉再生植株;分析银杉种源、培养基、植物生长调节剂等对银杉愈伤组织、不定芽、不定根诱导率和增殖壮芽率的影响。[结果]广西桂林种源银杉愈伤组织、不定根诱导率和增殖壮芽率最大,分别为66.48%,21.43%和55.00%。广西金秀种源银杉不定芽诱导率最大,为73.33%。适合银杉愈伤组织诱导植株再生的培养基和植物生长调节剂对于愈伤组织为DCR+0.5 mg·L^(−1)6-BA,对于不定芽为DCR+1.5 mg·L^(−1)6-BA,对于不定根为1/2 MS+0.2 mg·L^(−1)NAA+0.5 mg·L^(−1)IBA,对于增殖壮芽为DCR+1.0 mg·L^(−1)6-BA+0.4 mg·L^(−1)NAA。[结论]广西桂林种源银杉整体诱导率和增殖率相比广西金秀种源高。DCR培养基对银杉愈伤组织和不定芽诱导具有显著促进效果,1/2 MS培养基对不定根诱导具有显著促进效果。6-BA既可诱导银杉愈伤组织和又可诱导不定芽,NAA和IBA组合可诱导出银杉不定根,6-BA和NAA组合可对银杉不定芽进行增殖壮芽。

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。

Ezrin对小鼠卵母细胞成熟及早期胚胎发育的影响

旨在探究Ezrin对小鼠卵母细胞成熟、受精及早期胚胎发育的影响,揭示其可能作用机制。分别向小鼠GV期卵母细胞和原核期胚胎注射Ezrin的siRNA(si-Ez)或阴性对照siRNA(si-NC),观察对小鼠卵母细胞成熟、受精和早期胚胎发育的影响,并分别通过扫描电镜和免疫荧光染色的方法观察GV期注射si-Ez对小鼠成熟卵母细胞表面微绒毛生长、纺锤体定位和皮质颗粒迁移的影响。结果显示,GV期敲低Ezrin可显著降低小鼠卵母细胞成熟率,极显著降低受精后早期胚胎的囊胚率,虽可降低成熟卵母细胞的受精率,但差异不显著;在原核期敲低Ezrin,可极显著降低小鼠囊胚细胞数,桑椹胚存活率和囊胚率虽有下降趋势,但与对照组无显著差异;扫描电镜观察发现,敲低小鼠GV期卵母细胞Ezrin后显著降低了MⅡ期卵母细胞表面微绒毛的长度和密度;免疫荧光结果显示,敲低小鼠GV期卵母细胞Ezrin影响了纺锤体和皮质颗粒的迁移。结果表明,Ezrin通过影响微绒毛形成、纺锤体定位和皮质颗粒的正常迁移阻滞了小鼠卵母细胞成熟,进而影响了受精和早期胚胎发育。

水稻愈伤组织诱导及不定芽分化培养体系建立

目的:筛选高效诱导水稻愈伤组织及不定芽分化培养体系的培养基,为水稻基因工程育种奠定基础。方法:以优质水稻品种(润珠香占)成熟胚为外植体,研究成熟胚愈伤组织出愈情况、不同培养基以及植物生长物质浓度配比对愈伤组织诱导及不定芽分化的影响。结果:水稻愈伤诱导培养基出愈率大小为N6>B_(5)>MS>WPM,N_(6)与B_(5)、MS、WPM差异显著;2,4-D浓度愈伤组织诱导率大小为B_(4)>B_(3)>B_(5)>B_(6)>B_(2)>B_(1),B_(4)与各处理组之间差异显著;植物生长物质浓度配比对愈伤组织不定芽分化率大小为T_(5)>T_(7)>T_(6)>T_(1)>T_(8)>T_(3)>T_(2)>T_(4),T_(5)与各处理组之间差异显著。结论:水稻成熟胚愈伤诱导的最佳培养基及2,4-D质量浓度分别为N6、2.0 mg/L;不定芽分化最佳植物生长物质6-BA、KT、NAA质量浓度比为1∶1∶1,不定芽分化率最高。

LncRNA调控卵母细胞成熟及胚胎发育的作用机制研究进展

长链非编码RNA(lncRNA)在卵母细胞的发育和胚胎发育过程中发挥着重要作用。目前,lncRNA在卵母细胞成熟和胚胎发育中的具体作用机制尚不清楚,相关研究报道较少。本文综述了lncRNA调控卵母细胞成熟和胚胎发育的作用机制研究进展,着重讨论lncRNA在激素分泌、卵丘颗粒细胞、卵母细胞成熟以及合子基因组的激活等方面的调控作用,以期为进一步研究lncRNA提供新的启示。

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。