共查询到18条相似文献,搜索用时 93 毫秒
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本研究探讨了磷酸二酯酶(PDE)抑制剂Milrinone对水牛卵母细胞成熟的影响。水牛卵母细胞在含50μmol/L Milrinone的成熟液中培养16 h后,在不含Milrinone的成熟液中培养12 h、16 h和20 h,挑选形态正常的卵母细胞受精后培养,其分裂率(分别为59.1%,48.5%和50.6%)显著高于再培养8 h(29.5%)的卵母细胞(P0.05),但与其对照组(66.2%)相近,同时其卵母细胞受精后的囊胚发育率(分别为36.9%,33.6%和23.9%)亦显著高于8 h(15.5%)的卵母细胞(P0.05),但均与正常成熟培养的对照组(33.4%)相近。结果表明Milri-none对水牛卵母细胞的成熟具有明显的可逆抑制作用,经成熟抑制后的水牛卵母细胞恢复成熟后仍可保持较高的发育潜能。 相似文献
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表皮生长因子对水牛卵母细胞体外培养核质成熟的影响 总被引:1,自引:0,他引:1
为了探讨表皮生长因子(EGF)对水牛卵泡卵母细胞体外培养核质成熟的影响,在以TCM199为基础的成熟液中加入不同浓度的EGF(0、10、25、50、100 ng/ml),体外成熟培养24~26 h,观察第一极体(PB1)的排放;随后进行孤雌激活检测其分裂率、囊胚发育率、囊胚孵化率,并用Hoechst33342染色后计算囊胚的细胞数。结果发现添加EGF各组的卵母细胞第一极体排放率显著提高(P<0.05);成熟液中添加25 ng/ml EGF时,卵裂率及囊胚发育率(分别为80.0%、44.8%)明显高于对照组(分别为69.3%、31.9%,P<0.05),但对囊胚的细胞数影响不大。EGF不仅促进水牛卵母细胞体外培养的核成熟,而且有利于卵母细胞的胞质成熟,其中EGF的最佳浓度为25 ng/ml。 相似文献
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以皮层颗粒(CG)单层分布于质膜下做为卵母细胞胞质成熟的标志,利用CG荧光染色法,研究了不同浓度表皮生长因子(EGF)和胰岛素样生长因子(IGF-1)及其组合对水牛卵母细胞胞质成熟的影响。结果发现:(1)添加不同浓度的EGF(10、20、30、50ng/mL)都可以提高胞质的成熟率,但是组间差异不显著(P〉0.05);皮质颗粒的分布随着EGF浓度的升高逐步由中间分布向皮层分布转变;(2)在成熟液中添加IGF-130ng/mL时效果较好,能显著提高卵母细胞胞质的成熟率;皮质颗粒的分布随着IGF-1浓度的升高逐步由中间分布向皮层分布转变,在添加IGF一130ng/mL时,在皮层分布最好,随着IGF-1量的进一步增加,皮质颗粒又向中间分布转变;(3)添加20ng/mL EGF+30ng/mL IGF-1组卵母细胞体外成熟率高于添加30ng/mL的IGF-1组,显著高于添加20ng/m LEGF组和对照组(P〈0.05)。由此表明,EGF和IGF-1对水牛卵母细胞体外成熟有协同作用。 相似文献
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为了解卵母细胞体外成熟与凋亡过程,进而提高卵母细胞体外成熟率,本试验研究了在培养液中添加不同浓度的表皮生长因子(EGF)、胰岛素样生长因子-1(IGF-1)对水牛卵母细胞体外成熟和凋亡的影响。结果表明:(1)添加各种浓度的EGF(10,20,30,50,100 ng/mL)均可以提高水牛卵母细胞的成熟率,降低卵母细胞的凋亡率,其中50 ng/mL EGF有显著影响(P<0.05);(2)添加各种浓度的IGF-1(10,30,50,100 ng/mL)均能提高水牛卵母细胞体外成熟率,降低卵母细胞的凋亡率,以30 ng/mL效果明显(P<0.05);(3)添加20 ng/mL EGF+30ng/mL IGF-1组卵母细胞体外成熟率和凋亡率分别高于和低于单独添加IGF-1和EGF组。 相似文献
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主要探讨无卵丘水牛卵母细胞体外成熟的可行性,以便为研究卵母细胞成熟机理提供模型。无卵丘的水牛卵母细胞随机分为5组,然后分别进行直接成熟培养(M1),与卵丘细胞单层共培养(M2),用未扩展的卵丘细胞块包围培养(M3),与扩展的卵丘细胞团共培养(M4)和用卵巢组织包围培养(M5)。无卵丘的水牛卵母细胞体外成熟培养24 h后检查第一极体(PB1)排出率,随后对这些卵母细胞进行孤雌激活,评定其成熟质量。结果发现,M4组的第一极体排出率明显高于M1组和M5组,其它各组间没有显著差异(P〉0.05);M5组的孤雌激活卵裂率显著低于M1组和M4组(P〈0.05),而与M2组和M3组没有显著差异(P〉0.05),但M3和M4两组的囊胚发育率显著高于M1组和M5组(P〈0.05)。这些研究结果表明:(1)未扩展卵丘细胞包围法和扩展卵丘细胞团支撑法可促进无卵丘水牛卵母细胞的体外成熟,但与卵丘细胞单层共培养没有作用;(2)卵巢组织包围培养不利于水牛卵母细胞的体外成熟。 相似文献
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水牛与黄牛卵母细胞的回收及体外成熟、体外受精比较 总被引:6,自引:0,他引:6
在本文中 ,平均从每头水牛的卵巢上获得了 4 .5 8个卵母细胞 ,其中可用卵为 2 .98个 ,占 6 5 % ,卵母细胞在数量和质量上都低于黄牛 ,差异显著 (P <0 .0 1)。黄牛卵母细胞体外成熟、受精率显著高于水牛 (P <0 .0 5 )。 相似文献
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利用屠宰场屠宰的沼泽型水牛卵巢采集卵母细胞,统计了一年中4个季度1—3、4—6、7—9、10—12月份每个卵巢平均获卵母细胞总数、可用卵母细胞数及不可用卵母细胞数。结果:每个卵巢平均获卵母细胞总数分别为4.06枚±0.63枚、4.62枚±0.75枚、4.50枚±1.06枚、3.87枚±1.34枚,其中可用卵母细胞数分别为1.70枚±0.57枚、1.86枚±0.55枚、2.06枚±0.63枚和1.67枚±0.50枚;4个季度卵母细胞成熟率分别为:(81.5±6.2)%、(70.7±5.6)%、(70.7±9.3)%和(77.0±6.9)%。以上结果表明,在每个卵巢平均获卵母细胞总数上,4—6月份、7—9月份两季度显著高于10—12月份(P〈0.05),其他季节间没有显著性差异(P〉0.05);7—9月份每个卵巢平均获可用卵母细胞数最高,显著高于1—3月份和10—12月份(P〈0.05),但与4—6月份之间没有显著性差异(P〉0.05);4个季度不可用卵母细胞数之间没有差异(P〉0.05);4个季度卵母细胞的体外成熟能力上也没有显著差异(P〉0.05)。 相似文献
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Changes in the cytoplasmic inclusions during meiotic maturation were histochemically examined in cultured porcine oocytes. The oocytes contained a small amount of protein and glycogen granules throughout the maturation culture, as well as Sudanophilic lipids composed of small, medium and large droplets. Soon after collection, the amount of Sudanophilic lipid droplets of small and medium size was small and there were 167 ± 11.2 large droplets. After being cultured for 22 h, the number of large lipid droplets decreased remarkably, while the number of small and medium ones increased. There were no differences in the number of Sudanophilic lipid droplets of different sizes between ovulated oocytes and the oocytes cultured for 44 h. The oocytes always contained a large amount of neutral fats and lipoids, but not cholesterols. In the oocytes cultured for 22 h with olomoucine, both the resumption of nuclear maturation and the decrease in the size of the Sudanophilic lipid droplets were inhibited. From the present findings, it appears that the change in the size of the Sudanophilic lipid droplets in the cytoplasm of porcine oocytes is closely related to nuclear maturation. 相似文献
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Penglei Shen Jie Xu Ping Wang Xin Zhao Ben Huang Fei Wu Lulu Wang Weili Chen Yun Feng Zhenwei Guo Xiaohua Liu Yanfei Deng Jianrong Jiang Deshun Shi Fenghua Lu 《Reproduction in domestic animals》2020,55(2):170-180
At present, many three-dimensional (3D) culture systems have been reported, improving the oocyte quality of in vitro maturation (IVM), yet the mechanism still needs to be further explored. Here we examined the effects of a new self-made 3D glass scaffold on buffalo oocyte maturation; meanwhile, the underlying mechanism on buffalo oocyte maturation was also detected. Compared to the two-dimensional (2D) glass dish culture, results revealed that the 3D culture can improve the first polar body rate of oocytes, subsequent cleavage and blastocysts rate of parthenogenetic activation embryos (p < .05). The extracellular matrix-related proteins COL1A1, COL2A1, COL3A1, FN and cell connection-related proteins N-cadherin, E-cadherin, GJA1 were found higher in cumulus cells of 3D culture. Moreover, in cumulus cells, proteins of the PI3K/AKT pathway reported being regulated by FN and E-cadherin including PI3K P85 and p-AKT were also higher in 3D culture. Furthermore, proapoptosis proteins P53, BAX, caspase-3 were lower in both cumulus cells and oocytes in 3D culture, while proteins PCNA and BCL2 showed the opposite result. Results also showed that the apoptosis was inhibited, and the proliferation was enhanced in cumulus cells of 3D culture. Finally, the cumulus expansion-related genes HAS2, CD44, HMMR, PTX3, PTGS2 were found higher in cumulus cells of 3D culture. Taken together, the 3D culture could promote oocyte maturation by regulating proteins correlated with the ECM, cell connection and PI3K/AKT pathway, inhibiting the apoptosis of cumulus cells and oocytes, enhancing the proliferation of cumulus cells and the cumulus expansion. 相似文献
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Blasa Pereira Jesús Dorado Maria Diaz‐Jimenez Cesar Consuegra Isabel Ortiz Jaime Gosalvez Manuel Hidalgo 《Reproduction in domestic animals》2019,54(Z4):78-81
The acquisition of equine oocyte developmental capacity is ensured by the follicular environment, such as granulosa cells, which could reflect the meiotic development potential of immature oocytes. This study evaluated the relationship between DNA fragmentation of granulosa cells, using the chromatin dispersion test, and equine oocyte meiotic development after in vitro maturation. Granulosa cells and cumulus–oocytes complexes (n = 50) were recovered from slaughterhouse‐derived ovaries. Oocytes were in vitro matured, stained and evaluated under fluorescence microscopy. Maturation rates were classified into outstanding, medium and poor levels of maturation using 25th and 75th percentiles as thresholds. For DNA assessment, each sample was processed with the Ovoselect® kit (Halotech DNA). High, low and total DNA fragmentation percentages were compared among levels of maturation rates by ANOVA, followed by Duncan test. Results were expressed as mean ± SE. Total and high DNA fragmentation rates of granulosa cells were significantly higher (p < 0.05) in follicles whose oocytes had reached outstanding maturation level than those originating from follicles whose oocytes had reached poor maturation level. In conclusion, the DNA fragmentation analysis of equine granulosa cells can be a valuable test to identify equine oocytes showing the best meiotic competence after in vitro maturation. 相似文献
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Tatsanee PHERMTHAI Tamas SOMFAI Takashi NAGAI Rangsun PARNPAI 《Animal Science Journal》2012,83(9):630-638
The aim of the present study was to compare the efficiency of the solid surface (SSV), cryotop (CT) vitrification methods and cytochalasin B (CB) pretreatment for cryopreservation of immature buffalo oocytes. Cumulus‐oocyte complexes (COCs) were placed for 1 min in TCM199 containing 10% dimethylsulfoxide (DMSO), 10% ethylene glycol (EG), and 20% fetal bovine serum, and then transferred for 30 s to base medium containing 20% DMSO, 20% EG and 0.5 mol/L sucrose. CB pretreated ((+)CB) or non‐pretreated ((?)CB) COCs were vitrified either by SSV or CT. Surviving vitrified COCs were selected for in vitro maturation (IVM) and in vitro fertilization (IVF). The rate of viable oocytes after vitrification in CT groups (82%) was significantly lower (P < 0.05) than that in a fresh control group (100%), but significantly higher (P < 0.05) than those in SSV groups (71–72%). Among vitrified groups, the highest maturation rate was obtained in the CT (?)CB group (32%). After IVF, the cleavage and blastocyst formation rates were similar among vitrified groups but significantly lower than those of the control group. In conclusion, a higher survival rate of oocytes after vitrification and IVM was obtained in the CT group compared with that in the SSV group, indicating the superiority of the CT method. Pretreatment with CB did not increase the viability, maturation or embryo development of vitrified oocytes. 相似文献
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Vitrification by the Cryotop method is frequently used for bovine oocyte cryopreservation. Nevertheless, vitrified oocytes still have reduced developmental competency compared with fresh counterparts. The objective of this study was to compare the effect of vitrification either at the germinal vesicle (GV) stage or at the metaphase II (MII) stage on epigenetic characteristics of bovine oocytes and subsequently developing embryos. Our results demonstrated that vitrification of oocytes at each meiotic stage significantly reduced blastocyst development after in vitro fertilization (IVF). However, vitrification at the GV stage resulted in higher blastocyst development than did vitrification at the MII stage. Irrespective of the meiotic stage, oocyte vitrification did not affect 5-methylcytosine (5mC) immunostaining intensity in oocyte DNA. However, at both stages, it caused a similar reduction of 5mC levels in DNA of subsequently developing blastocysts. Oocyte vitrification had no effect on the intensity of H3K9me3 and acH3K9 immunostaining in oocytes and subsequent blastocysts. The results suggest that irrespective of meiotic stage, oocyte vitrification alters global methylation in resultant embryos although such alteration in the oocytes was not detected. Oocyte vitrification might not influence histone acetylation and methylation in oocytes and resultant embryos. Vitrification at the immature stage was more advantageous for blastocyst development than at the mature stage. 相似文献
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Jun Zhang Yanfei Deng Weili Chen Yonghong Zi Deshun Shi Fenghua Lu 《Reproduction in domestic animals》2020,55(11):1501-1510
Theca cells (TCs) play a key role in follicular growth and atresia. TCs synthesize androgens that act as substrate for granulosa cells (GCs) aromatization to estrogens needed for oocyte maturation. However, the effects of TCs in the form of conditioned medium on in vitro maturation (IVM) and developmental competence of buffalo oocytes remain unclear. In the present study, we examined the impacts of TC-conditioned medium (TCCM) on maturation efficiency and embryo development of buffalo oocytes after parthenogenic activation (PA). Our results showed that TCCM that was collected on day 2 and added to IVM medium at a 20% proportional level (2 days & 20%) exerted no significant effect on IVM rate (43.06% vs. 44.71%), but significantly (p < .05) enhanced embryo development (oocyte cleavage, 80.93% vs. 69.66%; blastocyst formation, 39.85% vs. 32.84%) of buffalo oocytes after PA compared with the control group. However, monolayer TC significantly (p < .05) promoted both maturation efficiency (48.84% vs. 44.53%) and embryo development (oocyte cleavage, 80.39% vs. 69.32%; blastocyst formation, 35.38% vs. 29.25%) of buffalo oocytes after PA compared to that in the control group. Furthermore, TCs secreted some testosterone into the conditioned medium, which significantly (p < .05) promoted the expression levels of oestrogen synthesis-related genes (CYP11A1, CYP19A1 and 17β-HSD) in buffalo cumulus–oocyte complexes (COCs). Our study indicated that TCCM (2 days & 20%) did not significantly affect IVM efficiency, but enhanced embryo developmental competence of oocytes after PA principally by stimulating the secretion of testosterone and facilitating estradiol synthesis of buffalo COCs. 相似文献