共查询到18条相似文献，搜索用时 250 毫秒
1.
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
2.
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
3.
研究了培养时间和激素对猪卵母细胞体外成熟的影响.结果表明:以NCSU-23为基础培养基的成熟培养液对卵母细胞分别培养36h、40h、44h、48h, 在培养44h后,成熟率最高为81.4%; 不同激素培养,成熟培养液培养22h后置于无激素培养液中继续培养22h的成熟率最高;添加性腺激素培养的成熟率显著高于不添加性腺激素的(P<0.05). 相似文献
4.
在卵母细胞体外成熟培养过程中,培养基中添加激素与否及其激素添加的先后顺序是影响猪卵母细胞核成熟和质成熟的一个重要因素.本试验将猪卵母细胞分别在FSH→不含激素、FSH→LH、FSH LH不含激素中培养48 h(培养第20～22 h后换液),并于成熟培养的第24 h(未换液)、48 h将卵母细胞进行荧光染色,观察其生发泡内染色质构型及卵母细胞核成熟情况.实验表明:(1)在IVM的前24 h,添加FSH LH组的GVIV期卵母细胞比例低于只添加FSH组,但差异不显著(8.99%比17.19%,P＞0.05);(2)在FSH存在的情况下,IVM的前期和后期添加LH能促进卵母细胞发生GVBD;(3)FSH LH培养24 h后转入不含激素培养基组,卵母细胞的核成熟比率显著高于添加FSH组和先添加FSH培养24 h后转入添加LH组(P＜0.05). 相似文献
5.
培养介质、卵丘细胞和卵泡直径对猪卵母细胞体外成熟的影响 总被引：29，自引：0，他引：29
A类卵母细胞在mTCM 199、NCSU2 3和NCSU37体系中培养 4 4～ 5 2小时后 ,成熟率分别为 76 .1%、78.1%和 6 5 .2 %。前两者差异不显著 (P >0 .0 5 ) ,但显著高于后者 (P <0 .0 5 )。卵母细胞在添加eCG和hCG的NCSU2 3体系中的成熟率 (75 .6 % )明显高于添加FSH的LH和成熟率 (6 5 .2 % ) (P <0 .0 5 )。A、B、C三类卵母细胞在NCSU2 3的成熟率分别为 73.3%、6 0 .4 %和 11.0 % ,三者间差异显著 (P <0 .0 5 )。大 (ф >6mm)、中 (ф =3～ 6mm)和小 (ф <3mm)三种卵泡中的卵母细胞在NCSU2 3中培养后 ,成熟率分别为 5 6 .2 % ,78.1%和 5 1.9% ,中等卵泡中卵母胞的体外成熟率显著高于其他两组 (P <0 .0 5 )。 相似文献
6.
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
7.
本研究就马尾藻属藻类多糖对猪卵母细胞的体外成熟及凋亡进行了探索：试验一证明，体外培养46h～48h时，成熟液添加马尾藻属藻类多糖能促进卵丘细胞扩展，添加20mg/ml马尾藻属藻类多糖能提高核成熟率(41．8%)，与对照组29．3%的核成熟率相比，二者差异显著；试验二证明，添加马尾藻属藻类多糖可延长猪卵母细胞体外存活时问，体外培养第16d，在含有0、5mg/ml、20mg/ml马尾藻属藻类多糖培养液中的猪卵母细胞存活率分别为2．6%、18．9%、23．0%，添加与否差异显著。 相似文献
8.
从延边黄牛卵巢中采集未成熟的卵母细胞进行体外成熟培养、体外受精及受精卵体外培养。结果表明 :1将卵子用 2种不同的培养液进行体外成熟培养和受精卵体外培养 ,TCM199组的卵裂率 (5 6 .3% )极显著高于 D- PBS组(33.6 % ) (P<0 .0 1) ;TCM199组的囊胚发育率和孵化率 (15 .1%、13.7% )虽高于 D- PBS组 (10 .5 %、8.7% ) ,但 2个组之间无显著差异 (P>0 .0 5 )。2以 TCM199作为基础培养液 ,分别用含激素培养液和不含激素培养液进行成熟培养和受精卵体外培养 ,添加激素组的卵裂率、囊胚发育率及囊胚孵化率 (81.2 %、17.5 %、15 .3% )高于没有添加激素的对照组 (75 .8%、12 .1%、10 .5 % ) ,但 2个组之间无显著差异 (P>0 .0 5 )。 3体外受精卵与单层颗粒细胞共培养组的卵裂率、囊胚发育率及囊胚孵化率 (78.0 %、11.5 %、9.9% )显著高于非共培养组 (6 8.1%、5 .4 %、3.6 % ) (P<0 .0 5 )。 相似文献
9.
本试验旨在探讨不同外源激素组合的添加、胰岛素—转铁蛋白—亚硒酸钠(ITS)的添加、不同培养皿、石蜡油的添加对卵母细胞体外成熟培养(IVM)的影响。结果显示:①孕马血清促性腺激素+人绒毛膜促性腺激素+促卵泡素(PMSG+HCG+FSH)组高于促卵泡素+促黄体素(FSH+LH)组和尿促性腺激素(hMG)组,相互之间差异显著(80.1%、68.3%、53.3%,P<0.05);②添加1% ITS到猪卵母细胞培养液中对卵母细胞成熟率无显著提高(P>0.05),但显著提高了孤雌激活后孤雌胚胎的卵裂率和囊胚率(63.3%、55.1%,18.7%、12.1%,P<0.05);③凹槽皿组猪卵母细胞成熟率显著高于四孔板和30 mm塑料皿组(73.3%、68.0%、68.3%,P<0.05);④石蜡油的添加对卵丘细胞扩展和卵母细胞体外成熟率均有显著提高(84.8%、69.9%,P<0.05)。结果表明培养液中添加PMSG+HCG+FSH和ITS及选用凹槽皿、成熟培养液上覆石蜡油可提高猪卵母细胞IVM效果。 相似文献
10.
培养液体积对牛卵母细胞体外成熟的影响 总被引：3，自引：0，他引：3
培养液体积影响牛卵母细胞的体外成熟 ,用四孔培养板 80 0 μL培养液与微滴 (5 0 μL)培养牛卵母细胞 ,其体外成熟率 (6 1.1%vs 6 2 .9% )和卵裂率 (34.4 %vs 35 .2 % )均无显著差异 (P >0 .0 5 ) ,但二者与不覆盖石蜡油的大体积培养液 (35mm× 10mm培养皿 2mL培养液 )中的各项指标 (成熟率 38.7%、卵裂率 2 3.2 % )差异均显著 (P <0 .0 5 )。 相似文献
11.
The aim of this study was to investigate the effects of culture in isolated oviducts relative to meiotic maturation, the time required to resume meiosis and the viability of the canine oocytes. For this purpose, cumulus–oocyte complexes and isthmus–ampullar tracts of the oviducts were collected from bitches undergoing ovariohysterectomies and destined to two experiments of culture. In experiment 1, the oocytes were cultured for 24 or 30 h: (1) in 100 μl drops under oil; (2) on the mucosal epithelium of the open oviducts; (3) in the ligated oviducts. In experiment 2, oocytes were cultured in the ligated oviduct for 24, 30 and 48 h. A group of control oocytes was not cultured (0 h). The results showed that within 30 h of culture, a higher proportion of oocytes (p < 0.001) resumed meiosis in the ligated oviduct (63.8%) than in drop (20.4%) or in the open oviduct (27.1%). Moreover, 24 and 30 h of culture assured higher proportions of meiosis resumption than 48 h (69.2 and 59.1% vs 35.8%, p < 0.005). Oocyte resumption of meiosis was mainly determined by oocytes at meiotic stages preceding metaphase I, while stages between metaphase I and II in the ligated oviduct ranged between 12.5 and 31.9%. The extension of the culture time up to 48 h in the oviduct increased oocyte degeneration significantly (59.3%, p < 0.0001) compared with 24 and 30 h (18.7 and 27.3%, respectively) and the oviductal epithelium showed nuclear picnosis and degeneration following culture. The present study suggests that the close physical interaction between the canine oocytes and the oviductal tract positively affects oocyte maturation, and meiosis is resumed within 30 h of culture. Moreover, the oocyte survival is better preserved within 30 h in the ligated oviduct compared with the conventional culture in drop or to the culture in the open oviduct, but the ligated oviduct does not assure viability of the oocytes up to 48 h of culture. 相似文献
12.
试验研究了清和促性腺激素对山羊卵丘扩展和卵母细胞核成熟的,卵母细胞与卵丘扩展的关系以及卵丘扩展与卵母细胞核成熟的关系。结果表明：（1）培养24h，添加PMSG组卵母细胞核成熟率显著高于不加激素组（P＜0．05），而培养到27h，2者成熟率之间差异变得不显著，说明添加PMSG卵母细胞核的最终成熟率没有明显影响，但加速了卵母细胞核的成熟进程；（2）M199＋BSA培养27h，卵母细胞核成熟率显著高于培养24h，而M199＋FCS培养27h与培养24h的成熟差异不显著，说明添加BSA时，卵母细胞核体外成熟速度比添加FCS的慢；（3）卵丘扩展良好与扩展不好的卵母细胞核成熟率以及第1极体形态无统计学差异，说明山羊卵母细胞的核成熟可能不依赖于卵丘扩展；（4）山羊的卵丘扩展产不依赖于卵母细胞；（5）将带壁颗粒细胞与不带壁颗粒细胞的COC分开培养发现，2者卵母细胞核成熟度无明显差异，带有壁颗粒细胞的COC卵丘扩展情况明显优于一般COC。 相似文献
13.
The aim of the present study was to improve cytoplasmic maturation of porcine oocytes by the addition of lycopene into in vitro maturation (IVM) media. We designed six experimental groups; IVM medium was supplemented with 10 IU/ml FSH, FSH and 10 IU/ml human chorionic gonadotrophin (hCG), or FSH and 7 μm lycopene in the first half of the IVM culture (0–22 h) followed by further culture (22–44 h) with or without hCG. The addition of lycopene into IVM media delayed the interruption of communication between an oocyte and the cumulus cells. Although meiotic competence was similar among the six groups, the glutathione level of matured oocytes was significantly higher in the lycopene‐supplemented group (9.89 pmol per oocyte) than that in other groups (7.25 and 7.81 pmol per oocyte). Fertilization rate was significantly improved in lycopene‐supplemented groups (58.3%) more than that in the group supplemented with FSH only (43.1%), whereas there were no differences in developmental competence among the groups (blastocyst rate: 20.1–29.5%). These results indicate that insufficient cytoplasmic maturation during conventional IVM resulted by disconnection of the gap junction between an oocyte and the cumulus cells in the early phase during IVM culture. We concluded that lycopene induced a prolonged sustainment of gap junctional communication between an oocyte and the cumulus cells during porcine IVM culture, which was an effective cytoplasmic maturation of porcine IVM oocytes. 相似文献
14.
15.
Tao Y Cao C Zhang M Fang F Liu Y Zhang Y Ding J Zhang X 《Journal of animal physiology and animal nutrition》2008,92(4):438-447
Cumulus cells (CCs) are of great importance in oocyte development and maturation in many species, but detailed influence of CCs has not been extensively examined, especially on rabbit. The present study was designed to investigate the effects of CCs and the elongation of in vitro maturation (IVM) time on rabbit oocyte nuclear and ooplasmic maturation and survival. Cumulus oocyte complexes (COCs) and naked oocytes (NOs) were recovered directly from rabbits super-ovulated with eCG. Corona-enclosed oocytes (COs) and denuded oocytes (DOs) were obtained from COCs after removing a part or whole of CCs. The oocytes were cultured in the following seven groups. (i) Cumulus cell enclosed oocytes (CEOs) were cultured alone (CEOs); (ii) COs were cultured alone (COs); (iii) DOs were cultured alone (DOs); (iv) NOs were cultured alone; (v) DOs were co-cultured with COCs [DOs(COCs)]; (vi) DOs were co-cultured with CCs [DOs(CCs)]; (vii) NOs were co-cultured with CCs [NOs(CCs)]. After the oocytes were cultured for 24 and 30 h, the nuclear maturation was evaluated by first polar body (PB1) extrusion while the ooplasmic maturation was evaluated by the cleavage rate after parthenogenetic activation. The results showed that the nuclear maturation rate of CEOs, COs, DOs(COCs) and DOs(CCs) after 24 h incubation were significantly different from each other (p < or = 0.05), the rate of DOs(CCs) was similar to that of DOs (p > or = 0.05). The cleavage rates in the first two groups were significantly higher than those of the others (p < 0.05). For oocytes cultured for 30 h, the nuclear maturation rates were significantly different for each culture model (p < 0.05). The cleavage rates in first two groups were significantly higher than those of others (p < 0.05). Both the nuclear and cleavage rates significantly increased when the culture time of DOs(COCs) was prolonged from 24 to 30 h. DOs(CCs) nuclear maturation was significantly improved when the culture time was prolonged from 24 to 30 h, but the ooplasmic maturation was not. Few NOs incubated with or without CCs accomplished nuclear maturation (approximately 2% both), even when the culture time was prolonged from 24 to 30 h. The oocyte degeneration rates were significantly different for each culture model after both 24 and 30 h incubation (p < or = 0.05). There was no significant difference in oocyte degeneration in the same groups between 24 and 30 h incubation (p > 0.05). The results suggest that rabbit CCs affect oocyte nuclear and ooplasmic maturation, and their survival. The prolongation of the culture time of rabbit oocyte from 24 to 30 h improves the nuclear and ooplasmic maturation differently in the present system. Rabbit oocytes free of CCs, especially NOs, show weak meiotic resumption potential and compromised viability, which cannot be improved by co-culture with dispersed CCs. The degeneration mostly happens at early time of IVM. 相似文献
16.
试验根据猪卵丘－卵母细胞复合体（COCs）的颗粒细胞层数,把COCs分成A、B、C和D级,A、B级用于体外分别培养24、32、38、44、48和54 h,C级用于体外分别培养24、32、38、44、48、54和60 h,观察它们不同时间排出极体数,然后将排出极体的卵母细胞孤雌激活。结果发现,A和B级COCs培养48 h时成熟率最高（83.2%和78.0%）,明显高于同级水平培养24、32和38 h的成熟率;不同级别COCs体外培养相同时间,A与B级成熟率差异不显著（P＞0.05）,但二者与C级的成熟率差异显著（P＜0.05）;体外培养48 h COCs卵裂率最高（81.7%）,与培养38 h前的卵裂率差异显著（P＜0.05）。结果表明,A、B级卵母细胞更适于体外培养,能获得高的成熟率和卵裂率,COCs周围颗粒细胞对卵母细胞的成熟有显著影响;COCs培养38 h之前,部分虽能看到极体,但激活后卵裂率极低,说明卵母细胞并未真正成熟,通常通过排出极体判断卵母细胞成熟是不够准确的,COCs体外培养44～48 h是成熟的最佳时期,能为体细胞核移植提供大量优质的MⅡ期卵母细胞。 相似文献
17.
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
18.
Chronological changes of bovine follicular cumulus-oocyte-complexesi were studied after in vitro maturation over a period of 48 h. According to their thickness and compactness of cumulus investments they were classified into 4 groups and cultured in enriched Ham’s F-10 medium with or without human chorionic gonadotrophin (hCG) and estradiolbenzoate (EB) for 0, 6, 12, 18, 21, 24, 27, 30 and 48 h. Representative samples were taken at each time interval for evaluation of nuclear maturation stages, ooplasm quality and size of the peri vitelline space (PVS). The results showed that oocyte nuclear breakdown (ONBD) required 6 to 12 h culture, and the peak of the first polar abstriction occurred at 24 h. The culture period required for ONBD and abstraction of the first polar body were related to the thickness and compactness of cumulus investments with and approximately 6 h delay in heavily compacted complexes. Ooplasm quality evaluation failed to show a clear trend, but the PVS increased in size from 0 h to 30 h and then, retracted again from 30 to 48 h. The overall maturation rate in the presence of hCG and EB was 79.1 %, and a substantial proportion (68.8 %) of nude or partially covered oocytes reached metaphase II stage. In the presence of hCG and EB no block at either metaphase I or at anaphase-telophase I was observed. In the absence of hCG and EB the percentage of oocytes reaching metaphase II was much lower (48.6%) in comparison with oocytes matured in the presence of these hormones (79.1 %). It was concluded a very high proportion of slaughterhouse oocytes could be matured in vitro and that the cumulus investments and addition of certain hormones affected the maturation rate. 相似文献