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1.
牛卵泡卵母细胞体外成熟的研究 总被引：3，自引：0，他引：3
研究了卵泡大小、卵泡液和卵丘细胞对牛卵泡卵母细胞体外成熟的影响。结果表明 :直径 2～ 6mm卵泡中的卵母细胞成熟率 ( 72 1%)最高 ,与直径小于 2mm( 5 8 4%)、6～ 8mm( 5 6 4%)及大于 8mm ( 3 5 0 %)卵泡中的卵母细胞组差异显著 ;成熟培养基中添加 10 %的新鲜牛卵泡液 (bFF)对牛卵母细胞的体外成熟有促进作用 ,但高浓度的bFF( 2 0 %、3 0 %)则抑制牛卵母细胞的体外成熟 ;卵丘 -卵母细胞复合体的质量影响卵母细胞的体外成熟 ,A、B、C三级卵母细胞成熟率分别为 86 1%、66 3 %、3 5 6%,卵裂率分别为 42 8%、3 1 9%、10 5 %,差异均显著 (P <0 0 5 )。 相似文献
2.
猪卵泡卵母细胞体外成熟与冷冻保存 总被引：1，自引：0，他引：1
研究了激素、猪卵泡液、不同类型血清对猪卵泡卵母细胞体外成熟的影响 ;比较了卵泡直径小于 2 mm、2～ 5 mm和大于 5 mm的卵母细胞体外成熟能力的差异 ,并对不同发育阶段猪卵母细胞的冷冻保存进行了研究。结果表明 :猪卵母细胞体外培养 4 8h时 ,培养的前 2 4 h培养液中加入激素 ,后 2 4 h去掉激素 ,卵母细胞的 A级成熟率 (5 1.73% )和总成熟率 (83.2 5 % )最高 ,极显著高于前 2 4 h不加激素 ,后 2 4 h添加激素培养的成熟率 (P<0 .0 1) ;也显著高于不含激素的培养液连续培养 4 8h的成熟率 (P<0 .0 5 ) ;但与添加激素连续培养 4 8h的成熟率差异不显著 (P>0 .0 5 )。在体外成熟培养液中 ,添加 10 % (体积分数 ) ECS的成熟率 (72 .86 % )显著高于添加 10 % (体积分数 ) NCS的成熟率(6 2 .2 1% ) (P<0 .0 5 ) ,而添加 10 % p FF则抑制卵母细胞的体外成熟。随着卵泡直径的增大 ,卵母细胞体外成熟能力逐渐增强。采用程序冷冻保存方法 ,成熟卵母细胞的成活率 (35 .5 9% )显著高于培养前 (2 4 .6 4 % )和培养 2 4 h(2 3.36 % ) (P<0 .0 5 )。培养 2 4 h(77.2 2 % )和培养成熟 (72 .81% )的卵母细胞解冻后的形态完整率均显著高于培养前(5 3.2 4 % ) (P<0 .0 5 ) 相似文献
3.
4.
猪小腔卵泡卵母细胞体外成熟研究 总被引：27，自引：0，他引：27
本实验通过与2～6mm猪卵泡卵母细胞相对比，研究了猪2mm以下卵泡卵母细胞的体外成熟。结果如下(1)由2mm以下卵泡获得的小(直径100.29±7.9μm)、中(109.36±3.71μm)和大(118.90±2.46μm)3类卵母细胞培养48h的成熟率分别为0、22.31％和45.45％。(2)2mm以下卵泡卵母细胞的成熟率(52.63％)显著低于2～6mm卵泡卵母细胞的成熟率(80.39％)。(3)以含PFF的培养液培养2mm以下卵泡卵母细胞其成熟率(53.54％)显著高于不含PFF的对照组(22.77％)。(4)在培养的前24h加入PMSG、后24h去除PMSG，其成熟率(41.86％)与含PMSG的培养液连续培养48h的成熟率(50.0％)无显著差异，而显著高于前24h不加PMSG，后24h加入PMSG培养的成熟率(5.41％)；也显著高于以下不含PMSG的培养液连续培养48h的成熟率(3.3％)。 相似文献
5.
PMSG和hCG对猪卵母细胞体外成熟的影响 总被引：20，自引：2，他引：18
对猪卵母细胞不同发育阶段的激素需要进行了初步探讨。结果表明 :猪卵母细胞体外培养 48h ,前 2 4h在培养液中加入激素 ,后 2 4h不加激素 ,卵母细胞的A级成熟率 (51 73% )和总成熟率 (83 2 5 % )最高 ,极显著高于前 2 4h不加激素 ,后 2 4h添加激素培养的成熟率 (P <0 0 1 ) ;也显著高于不含激素的培养液连续培养 48h的成熟率 (P <0 0 5) ;但与添加激素连续培养 48h组成熟率差异不显著 (P >0 0 5)。 相似文献
6.
马卵母细胞胞质内精子注射后体外发育能力的研究 总被引：2，自引：0，他引：2
本研究在非繁殖季节评估卵丘形态(松散型、致密型)、成熟培养体系(TCM 199、NCSU 23)、体外成熟时间(34、38 h)和离子霉素结合6-DMAP激活对马卵母细胞胞质内精子注射(ICSI)后体外发育能力的影响。从屠宰场采集马卵巢,获得的卵母细胞进行体外成熟,然后注射马冷冻解冻精液,统计分裂情况。试验结果表明,①马松散型卵母细胞成熟率显著高于致密型卵母细胞(P<0.05),分别为61.09%和41.24%,但ICSI后36 h分裂率无显著差异(P>0.05),分别为47.34%和44.92%;②两种培养体系对马松散型或致密型卵母细胞成熟率及ICSI后36 h分裂率无显著影响(P>0.05),但相同成熟体系培养松散型卵母细胞成熟率均显著高于致密型卵母细胞(P<0.05),然而ICSI后36 h分裂率差异不显著(P>0.05);③松散型或致密型卵母细胞在TCM 199或NCSU 23中成熟38 h成熟率均高于34 h成熟率,分别为44.43%～68.87%和34.52%～58.90%,松散型卵母细胞在TCM 199体系中成熟34 h、ICSI后激活组或对照组的分裂率显著高于成熟38 h、ICSI后激活组的分裂率(P<0.05),以及致密型卵母细胞在TCM 199体系中成熟34 h、ICSI后激活组的分裂率(P<0.05),而且显著高于松散型卵母细胞在NCSU 23体系中成熟38 h、ICSI后对照组的分裂率(P<0.05);④ICSI后用离子霉素结合6-DMAP激活对马卵母细胞ICSI后36 h分裂无显著影响(P>0.05)。因此,马松散型和致密型卵母细胞的成熟能力存在差异,TCM 199和NCSU 23成熟体系对这2种类型卵母细胞的发育能力无显著影响(P>0.05),马卵母细胞成熟38 h成熟率高于34 h成熟率,TCM 199成熟体系培养松散型卵母细胞34 h进行ICSI后的分裂率最高。离子霉素结合6-DMAP激活对TCM 199或NCSU 23体系成熟马卵母细胞ICSI后的体外发育能力无显著影响(P>0.05)。 相似文献
7.
在成熟过程中添加牛血清和猪卵泡液对猪卵母细胞核成熟及体外受精后早期胚胎发育的影响 总被引：24，自引：0，他引：24
研究目的在于探讨在成熟过程中添加牛血清和猪卵泡液对猪卵母细胞核成熟、卵丘细胞扩散及体外受精后早期胚胎发育的影响。卵母细胞·卵丘细胞复合体在含FSH和LH的以下处理组的成熟液中成熟培养 2 3～ 2 4h :(1)对照组-改良TCM - 199+0 .1%PVA ;(2 )试验组 1-改良TCM - 199+10 %新生牛血清 ;(3)试验组 2 -改良TCM - 199+10 %猪卵泡液 ,再移至无FSH和LH的不同处理组的成熟液中成熟培养 2 3～ 34h。试验 1中 ,卵母细胞在 4 6～ 4 8h成熟培养后 ,观察卵丘细胞扩散情况 ,并对卵母细胞进行固定和染色 ,鉴定卵母细胞减数分裂情况 :试验 2中 ,对在不同处理组的成熟液中成熟培养 4 6～ 4 8h的卵母细胞进行体外受精 ,再培养 8d。受精后第 2天检查分裂率、第 6天检查桑椹胚 /囊胚率、第 8天检查囊胚率。 4 6～ 4 8h成熟培养后试验组 1和试验组 2的大部分卵母细胞 -卵丘细胞复合体的卵丘细胞完全扩散 ,而对照组的卵丘细胞只有 5 0 %扩散。试验组 1和试验组 2的卵母细胞核成熟率分别为 39.9% (77/ 193)和 4 4 .3% (93/ 2 10 ) ,与对照组的卵母细胞核成熟率 4 8.1% (99/ 2 0 6 )相比没有显著差异 (P <0 .0 5 )。卵母细胞分裂率试验组 1(5 0 .0± 1.8) %和试验组 2 (49.9± 2 .6 ) %与对照组的卵母细胞分裂率 (49.0± 2 相似文献
8.
试验利用水牛卵泡液(BuFF)和黄牛卵泡液(BoFF)对不同来源水牛卵母细胞体外受精效果的影响进行了探讨,以完善水牛体外受精培养系统,进一步提高水牛胚胎体外生产效率。试验按成熟培养液中添加卵泡液替代胎牛血清量共分4个组。不添加卵泡液(0%+10%胎牛血清)为Ⅰ组(对照组);添加5%卵泡液+5%胎牛血清为Ⅱ组;添加10%卵泡液+0%胎牛血清为Ⅲ组;添加15%卵泡液+0%胎牛血清为Ⅳ组。结果表明,添加BuFF对活体采集卵母细胞和屠宰场收集卵母细胞的体外受精卵分裂率无显著影响(P0.05),但添加5%和10%BuFF对卵母细胞体外受精后的胚胎发育有明显促进作用,囊胚率均极显著高于对照组和15%BuFF组(P0.01),5%和10%BuFF组间无显著差异(P0.05);添加15%BuFF囊胚率有降低的趋势,但与对照组相比差异不显著(P0.05)。而添加10%BoFF组活体采集卵母细胞体外受精的受精卵分裂率和囊胚率均极显著高于对照组和5%BoFF组(P0.01),添加5%BoFF组的受精卵分裂率和囊胚率与对照组无显著差异(P0.05);添加BoFF对屠宰场收集水牛卵母细胞体外受精卵分裂率无显著差异(P0.05),但添加5%和10%BoFF组的囊胚率均显著高于对照组和15%组(P0.05),添加15%BoFF组与对照组相比,囊胚率显著降低(P0.05),5%和10%BoFF组间囊胚率无显著差异(P0.05)。综合以上结果,在水牛卵母细胞成熟培养液中添加5%～10%的BuFF或BoFF代替牛血清,可明显提高水牛体外胚胎生产效率,且以添加同种的BuFF效果略好。 相似文献
9.
用切割法采集卵泡液,收集卵丘一卵母细胞复合体（Cumulus oocytes comlexs,COCs）和自然裸卵,将部分COCs去除卵丘细胞获得机械裸卵,COCs放入体外成熟培养液中培养为成熟卵母细胞,加入获能的精子液,进行体外受精。结果表明：卵母细胞的体外成熟率和卵裂率与卵泡直径密切相关,大卵泡（80．95％,P〈0．01）和中等卵泡（75．50％,P〈0．05）的卵母细胞成熟率高于小卵泡（50．27％）;犬卯泡（53．53％）和中等卵泡（47．13％）的卵裂率显著高于小卵泡的32．26％（P〈0．05）。COCs、机械裸卵和自然裸卵的体外成熟率分别为75．0％、54．2％和10．5％,差异极显著（P〈0．01）,卵裂率分别为53．8％、10．8％和0％,差异极显著（P〈0．01）。对照组和1×10^5、1×10^6个／mL颗粒细胞组卵母细胞体外成熟率分别为68．6％、69．6％和67．8％,无显著差异（P〉0．05）,但均显著高于1×10^7个／mL（51．5％,P〈0．05）和1×10^10个／mL（35．5％,P〈0．05）颗粒细胞组,但各组间的体外受精率无显著差异（P〉0．05）。结果提示,大卵泡和中卵泡的卵母细胞的体外成熟率和卵裂率显著高于小卵泡,体外成熟培养液中添加高浓度的颗粒细胞能显著抑制卵母细胞的体外成熟。 相似文献
10.
体外成熟对牛卵母细胞孤雌激活后发育潜力的影响 总被引：2，自引：1，他引：1
本试验在卵母细胞体外成熟的研究基础上,研究了培养用水、卵泡液和成熟时间对卵母细胞体外成熟及孤雌激活后发育潜力的影响.结果表明(1)将卵母细胞分别于蒸馏水和Milli-Q超纯水配制的成熟培养液中培养22～24 h, 孤雌激活后, 卵裂率无显著差异(P>0.05); 但孤雌卵在超纯水配制的胚胎培养液中培养,囊胚发育率为22.3%, 明显高于在蒸馏水配制的培养液中的囊胚发育率14.1%(P<0.05).(2)在成熟培养液中添加10%、20%卵泡液,成熟卵母细胞孤雌激活后的囊胚发育率(32.3%, 30.9%)明显高于添加5%和0%卵泡液组的囊胚发育率(21.8%, 22.7%,P<0.05).卵母细胞在添加20%卵泡液的培养液中成熟培养,孤雌激活后囊胚孵化率明显低于添加0、5%、10%卵泡液组的囊胚孵化率(P<0.05).(3)成熟28 h或30 h的卵母细胞孤雌激活后,其囊胚发育率(30.5%或29.4%)明显高于成熟24 h或26 h组的囊胚发育率(20.5%或22.1%,P<0.05). 相似文献
11.
Meiotic Competence of Canine Oocytes Embedded in Collagen Gel 总被引：1，自引：0，他引：1
T Otoi R Shimizu H Naoi P Wongsrikeao B Agung M Taniguchi 《Reproduction in domestic animals》2006,41(1):17-21
The present study was conducted to examine the meiotic competence of canine oocytes embedded in collagen gel, and to investigate the effects of timed exposure of the oocytes embedded in collagen gel to gonadotrophins during maturation culture, on their nuclear maturation. Cumulus-oocyte complexes (COCs) were collected from bitches at the anoestrous and dioestrous stages of the reproductive cycle. In the first experiment, half of the COCs were embedded in collagen gels. The COCs with or without collagen-gel embedding were cultured in a TCM-199 medium supplemented with 0.1 IU/ml human menopausal gonadotropin (hMG) and 10 IU/ml human chorionic gonadotropin (hCG) for 72 h. In the second experiment, the COCs embedded in collagen gels were cultured in TCM-199 medium with gonadotrophins (hMG and hCG) for various periods (0, 24, 48 and 72 h) and then cultured in the medium without gonadotrophins until reaching total culture period (72 h). The percentage of the oocytes reaching metaphase I and metaphase II (MI/MII) was significantly higher (p < 0.05) in COCs with collagen-gel embedding than in COCs without collagen-gel embedding. The percentage of oocytes that were arrested at the germinal vesicle stage was significantly lower (p < 0.05) in oocytes cultured with gonadotrophins than in oocytes cultured without gonadotrophins. However, there were no significant differences in the percentages of oocytes that reached each stage of meiosis among the groups, irrespective of the duration of exposure to gonadotrophins. These observations indicate that embedding of COCs by collagen gel enhances the meiotic competence of canine oocytes, but removal of hormone supplement from maturation medium does not improve the ability of the oocytes to reach MII stage. 相似文献
12.
The effect of preservation condition of ovaries on the in vitro maturation of the porcine oocytes was studied. Cumulus‐oocyte complexes (COCs) were obtained from the ovaries preserved in Dulbecco’s phosphate buffered saline (PBS) solution at various temperatures for different time intervals, and cultured in M199 maturation medium. Matured oocytes were obtained from the ovaries preserved in PBS for 8 h and electrically activated. The activated oocytes were then cultured in NCSU23 embryo culture medium for 16 h to observe activation or 144 h to observe embryo development. It was found that the preservation temperature affected the maturation of porcine oocytes greatly. The effect was described as a compromise of the suppressions of autolysis at physiological temperature and frostbite because of low temperature. A preservation temperature of approximately 25°C showed the maximum maturation rate for a preservation time of 8 h. Preservation temperature also affected the activation and embryo development of porcine oocytes greatly, following a trend similar to the effect of preservation temperature on the maturation. Based on maturation rate, activation rate and cleavage rate, a preservation temperature of approximately 25°C would be optimum for a preservation time of 8 h. 相似文献
13.
Meiotic Response of In vitro Matured Canine Oocytes under Different Proteins and Heterologous Hormone Supplementation 总被引：2，自引：0，他引：2
The impact of TCM‐199 supplemented with different proteins and heterologous hormones on the in vitro maturation (IVM) rate of bitch oocytes was evaluated by nuclear staining under fluorescence microscopy. Oocytes were recovered by slicing of ovaries from bitches presented at various stages of oestrous cycle to ovariohysterectomy. The basic culture medium was TCM‐199 supplemented with 25 mM Hepes/l, with 10% heat‐inactivated oestrous cow serum (ECS), 50 μg/ml gentamicin, 2.2 mg/ml sodium bicarbonate and 22‐μg/ml pyruvic acid, 1.0‐μg/ml oestradiol (E 8875; Sigma), 0.5‐μg/ml follicle‐stimulating hormone (FSH) (Folltropin‐V; Vetrepharm Inc., Ontario, Canada) and 0.03 IU/ml human gonadotropin (hCG) (Profasi HP; Serono, Aubonne, Switzerland). Oocytes were distributed randomly between basic culture medium (control) and the corresponding experimental treatment. Hormone treatments were: oocytes cultured in; (1) medium without FSH, (2) control medium supplemented with 20 μg/ml oestradiol, or (3) medium supplemented with 1 μg/ml human somatotropin (hST; Humatrope, Lilly, Saint Cloud, France). The second experiment consisted of oocytes cultured in medium supplemented with 0.4% (w/v) bovine serum albumin (BSA, fraction V; Gibco Grand Island, NY, USA) instead of ECS, or oocytes cultured in medium with 10% inactivated oestrous bitch serum (EBS) instead of ECS. Oocytes were cultured in 100 μl droplets (up to 25 oocytes per drop) under mineral oil at 37°C in a 100% humidified atmosphere containing 5% CO2 in air. After 72 h of IVM, the highest rates (p < 0.05) of meiotic resumption were achieved with the 0.4% BSA supplementation. A positive influence on the metaphase II (MII) acquisition rate was observed with hST supplement. Oocytes cultured with 10% EBS supplementation did not develop to the MII stage. The results in this study show that the protein and hormone supplements to TCM‐199 culture medium tested did not promote the final steps of IVM of bitch oocytes. 相似文献
14.
Ahmed Sabry S. Abdoon Hassan A. Abdel-Rahman Sherif M. Shawki Omaima M. Kandil Said I. Fathalla 《Veterinary research communications》2014,38(4):287-295
This work was designed to evaluate the ovarian follicular development, oocytes morphology, methods of oocytes reterival, and the effect of different in vitro maturation (IVM) media on cumulus cell expansion and nuclear maturation of Jennies oocytes. Experiment 1, the number of small (<6 mm), medium (6 to 9 mm) and large size (>10 mm) ovarian follicles was recorded. Cumulus-oocyte-complexes (COCs) were reterived and classified into 4 Grades based on their cumulus-cells investment and the homogenous of the ooplasm. In Experiment 2, COCs were recovered by using 18-G, 20-G needle or slicing and scraping of ovarian follicles to determine the number and morphology of the recovered COCs. In Experiment 3, Grade A and B COCs were IVM in DMEM-HG, DMEM-LG, DMEM-F12, TCM199, TCM199-F12 or CR1aa media supplemented with 10 % FCS?+?10 μg FSH/mL?+?10 IU hCG/mL?+?50 μg/mL gentamicin. Maturation was performed for 36 h at 38.5 °C under 5 % CO2 in humidified air. After IVM, cumulus cell expansion and oocytes nuclear canfiguration were determined. An average of 6.40?±?0.26 follicles was recorded per Jenny ovary, representing 3.37?±?0.46, 1.89?±?0.14 and 1.14?±?0.16, for the small, medium and large size follicles, respectively. Oocyte recovery was higher (P?P?P?P?P?P?P?P?Conclusion: Slicing and scraping or aspiration of follicles using 18-G needle increased the number and percentage of Grade A Jennies oocytes. TCM199-F12, CR1aa and TCM199 medi are more suitable for IVM of Jenny oocytes by promoting cumulus cells expansion and nuclear maturation to M II stage. 相似文献
15.
The objectives of the present study were to investigate the relationship between the morphological status of cumulus cells surrounding canine oocytes after maturation culture and the meiotic stage of the oocytes. In addition, the effect of the removal of cumulus cells from canine cumulus-oocyte complexes (COCs) during maturation culture on their meiotic competence was examined. Canine COCs were collected from bitches at the anoestrous and dioestrous stages and only COCs with >110 microm in vitelline diameter were cultured in medium 199 with 10% canine serum for 72 h. In the first experiment, the relation between the morphological status of cumulus cells surrounding oocytes cultured for 72 h and their meiotic stages was examined. At the end of maturation culture, the proportions of intact, partially nude and completely nude oocytes were 65.2%, 22.9% and 11.9%, respectively. The proportion of maturation to metaphase II of completely nude oocytes was highest among the oocytes with different morphological status of cumulus cells. In the second experiment, the cumulus cells were partially or completely removed from COCs at 48 h after the start of maturation culture and the oocytes were cultured for a further 24 h. The proportion of oocytes reaching metaphase II in the completely denuded oocytes was significantly higher than that in the control oocytes without the removal treatment of cumulus cells. The results indicate that morphological status of cumulus cells surrounding oocytes may be related to the nuclear maturation of canine oocytes, and the removal of cumulus cells from COCs during maturation culture can promote the completion of oocyte meiotic maturation. 相似文献
16.
Predictive value of the area of expanded cumulus mass on development of porcine oocytes matured and fertilized in vitro 总被引：3，自引：0，他引：3
The objective of the present study was to investigate the correlation between the degree of cumulus expansion and in vitro development of porcine cumulus-oocytes complexes (COCs) matured and fertilized in vitro. The COCs were matured in the maturation medium (IVMM) supplemented with 15% or 5% of porcine follicular fluid (PFF) from small, medium and large follicles (<2 mm, 2-5 mm and >5 mm, respectively). COCs cultured in IVMM with PFF for 48 h displayed less expansion than those cultured in IVMM alone (P<0.05), irrespective of follicle size. After culture for 24 h in IVMM with PFF and for another 24 h in IVMM alone, the degree of cumulus expansion was more prominent than culture in the presence of PFF for the entire 48 h period (P<0.05), but the percentages of oocytes with PB I showed no significant difference between the control and experimental groups (P>0.05). After in vitro fertilization, the oocytes failed to develop to the morula/blastocyst stages except for those matured in IVMM supplemented with 15% or 5% PFF obtained from >5 mm follicles for the first 24 h and followed by in IVMM alone for the second 24 h (12.5% and 11.1% of the embryos developed to morulae and blastocysts, respectively). The expanded cumulus areas of COCs were significantly positively correlated with their in vitro development (p=0.0058, 0.0001 and 0.0348 for the percentages of embryos developed to 2-4 cell, beyond 4 cell and morula and blastocyst stages, respectively). In conclusion, PFF had an inhibiting effect on cumulus expansion, and the inhibitory effect decreased progressively with the increase in size of follicles from which PFF was obtained, and the action of PFF on cumulus expansion was affected by the PFF culture time. The areas of the expanded cumulus mass may be used as a parameter to predict development of porcine oocytes matured and fertilized in vitro. 相似文献
17.
Effect of Follicle Size on In Vitro Maturation of Pre‐Pubertal Porcine Cumulus Oocyte Complexes 
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下载免费PDF全文 Very small follicles (<3.0 mm diameter) are over‐represented on the surface of ovaries of non‐cycling pigs, and the oocytes collected from these follicles generally have reduced developmental competence in vitro. This study examined the effect of follicle size on the nuclear maturation (n = 608), the potential of parthenogenetic activation (n = 243) and the cyclic AMP (cAMP) content of pre‐pubertal porcine oocytes (n = 480). In addition, the influence of follicle size on steroid hormone synthesis was analysed. Cumulus oocyte complexes (COCs) flushed from small (2.5–4.0 mm) or large (4.5–6.0 mm) ovarian follicles were cultured for 0, 28 and 46 h. After 46 h of IVM, a greater proportion of oocytes from 4.5‐ to 6.0‐mm follicles reach metaphase II (MII) compared with those from follicles with 2.5–4.0 mm of diameter (96.1 vs 77.0%, respectively; p < 0.001). Parthenogenetic activation of oocytes from large follicles produced higher developmental rates than oocytes from large follicles (p < 0.05). At 28 h, the IVM medium with oocytes from large follicles contained significantly more 17ß‐oestradiol (E2) than the medium with oocytes from small follicles (5.55 vs 3.45 ng/ml, respectively; p < 0.05) and at 46 h, the medium with oocytes from small follicles contained significantly more progesterone (P4) than the medium with oocytes from large follicles (276.7 vs 108.2 ng/ml, respectively, p < 0.05). Porcine oocytes from large follicles have higher nuclear and cytoplasmic maturation capacities, but the differences did not appear to be cAMP‐mediated. Our findings also suggest that COCs from small follicles undergo more intensive luteinization than COCs from large follicles. The results show that oocytes from follicles with a diameter greater than 4.0 mm are more suitable for in vitro studies. 相似文献
18.
Effects of bovine serum albumin and estrous cow serum on development and ultrastructure of in vitro-produced porcine embryos 总被引：1，自引：0，他引：1
This study evaluated the effects of bovine serum albumin (BSA; 4 mg/ml) and estrous cow serum (ECS; 10%) in North Carolina State University (NCSU) 23 medium on the development of in vitro-matured and in vitro-fertilized porcine oocytes. Early cleavage rate was significantly (P < 0.05) higher in NCSU/ECS (71.3 +/- 14.7%) vs. NCSU/BSA (60.6 +/- 4.7%). Cleavage beyond the four-cell stage was not different between the two culture media (43.5 +/- 9.5% and 41.4 +/- 17.7%, respectively). The proportion of development to blastocysts was--with borderline significance (P = 0.05)--higher in NCSU/BSA (28.0 +/- 4.4%) than in NCSU/ECS (20.4 +/- 7.3%). Blastocysts produced in NCSU/BSA had significantly (P < 0.001) higher cell numbers than those cultured in NCSU/ECS (29.5 +/- 20.1 vs. 16.9 +/- 10.8). The ultrastructure of in vitro-produced blastocysts from both culture systems was compared vs. in vivo-derived blastocysts. The latter showed a clear differentiation between trophectoderm (TE) and inner cell mass (ICM) cells. The TE cells were anchored to other TE cells or ICM cells by long, well-developed junctional complexes. The apical membrane of trophoblast cells was covered with numerous microvilli. Mitochondria were abundant, round to elongated in shape, and showed clear transverse cristae. The ultrastructure of blastocysts cultured in NCSU/BSA mimicked that of in vivo-derived embryos closely. In contrast, blastocysts from the NCSU/ECS culture system displayed an irregular ultrastructure with reduced numbers of organelles and numerous cytoplasmic inclusions, such as lipid-yolk-vacuoles and vacuoles with lipid content. In some sections of these embryos, cellular debris was detected in cytoplasm. The shape of mitochondria was more ovoid and cristae were not visible. In summary, our results demonstrate a beneficial influence of ECS in the culture medium on initial cleavage of in vitro-produced porcine embryos. Clearly negative effects of ECS in the subsequent culture period are associated with marked ultrastructural changes of embryonic cells. 相似文献