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以屠宰场牦牛卵巢为材料,比较了抽吸加切割法和抽吸法2种卵母细胞离体采集方法的效率和5种成熟培养液的培养效果,并研究了卵泡位置、形态对卵母细胞体外成熟的影响。结果表明:在牦牛乏情期,平均每个卵巢用抽吸加切割法回收卵数极显著高于抽吸法(9.33±4.30VS4.70±2.62,P<0.01),可用卵数也极显著高于抽吸法(5.63±4.19VS4.37±2.32,P<0.01)。将牦牛卵丘卵母细胞复合体(COCs)分别置于5种成熟培养液中培养,其中M199(缓冲体系为Earles盐) 10% FBS 5.0mg/LLH 1.0mg/LE2 双抗(青霉素100IU/mL和链霉素100mg/L)的成熟液效果最好,成熟率为81.33%,卵裂率为49.33%。来自卵巢表面卵泡的COCs的成熟率和卵裂率均高于来自卵巢内卵泡的COCs(分别为81.33%VS69.33%,P>0.05;49.33%VS34.67%,P<0.05)。来自明亮卵泡的COCs的成熟率和卵裂率均极显著高于来自浑浊卵泡的COCs(分别为81.33%VS33.33%,P<0.01;49.33%VS3.33%,P<0.01)。 相似文献
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Lange Consiglio A. Bignotti A. Pecile A. M. Cremonesi F. 《Veterinary research communications》2009,33(1):89-92
Veterinary Research Communications - 相似文献
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牦牛卵巢卵母细胞体外培养成熟条件的建立 总被引：2，自引：2，他引：2
研究了不同采集方法和不同培养液对牦牛卵母细胞体外成熟的培养效果。结果:在牦牛乏情期,每个卵巢平均回收卵数为(9.33±4.30),可用卵数为(5.63±4.19)。将牦牛卵丘卵母细胞复合体分别置于5种成熟培养液中培养,成熟率分别为75.56%、71.11%、81.33%、77.33%和77.78%,卵裂率分别为42.22%、33.33%、49.33%、46.67%和42.22%,其中C的成熟液效果最好。来自卵巢表面卵泡的COCs的成熟率(81.33(vs69.33(,P>0.05)高于来自卵巢内卵泡的COCs,卵裂率(49.33%vs34.67%,P<0.05)显著高于来自卵巢内卵泡的COCs。来自明亮卵泡的COCs的成熟率(81.33%vs33.33%,P<0.01)和卵裂率(49.33%vs3.33%,P<0.01)极显著高于来自混浊卵泡的COCs。 相似文献
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Recent improvements in cryopreservation of mammalian eggs enable the long-term preservation of female germ cells in several mammalian species. Nevertheless, cryopreservation of porcine oocytes is still considered as a challenge. Although the use of vitrification techniques result in reasonable survival rates, developmental competence of vitrified oocytes has been compromised. Alterations of zona characteristics, cytoskeleton, mitochondrial functions and antioxidant-defense ability caused by vitrification are among the most frequently observed malformations which may be responsible for the low developmental competence of cryopreserved porcine oocytes. Furthermore, in vitro maturation, fertilization and embryo culture technologies, which are indispensable for generating embryos from cryopreserved oocytes, generate high rates of abnormal fertilization (polyspermy) and additional stress in resultant embryos further compromising their developmental competence. As a result, embryo development of porcine cryopreserved oocytes is still at low level and to date no piglet has been produced from such oocytes. The aim of the present review is to summarize knowledge on viability and developmental competence of vitrified porcine oocytes and to give ideas for future perspectives for the improvement of porcine oocyte cryopreservation technology. 相似文献
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牛卵母细胞的体外成熟 总被引：5，自引：1，他引：4
在卵母细胞体外成熟液（含FSH）中分别添加10、30、50μg/L表皮生长因子（EGF）,24 h检查成熟率。结果表明,添加10、30μg/L EGF组牛卵母细胞成熟率为79.8%、71.5%与对照组（未添加EGF）成熟率（70.4%）无明显差异（P〉0.05）,但EGF添加至50μg/L时牛卵母细胞第一极体（PB1）排出率显著提高,达85.4%（P〈0.01）;在此基础上,比较了成熟液中添加尿促性素（HMG）对牛卵母细胞体外成熟的影响,结果发现在含FSH和EGF的成熟液中添加HMG未能显著提高牛卵母细胞体外成熟效果（P〉0.05）;最后,比较了HMG对牛卵母细胞体外成熟的效果,结果发现单独添加HMG成熟效果显著高于FSH（P〈0.05）,表明成熟液中如不含其他生长因子,则单独添加HMG较FSH对牛卵母细胞体外成熟的效果好。 相似文献
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采用PBS作为冷冻基础液,分别用甘油和二甲基亚砜(DMSO)作为冷冻保护液,在程序化冷冻保存和玻璃化冷冻保存条件下,研究小鼠生发泡期(GV期)卵母细胞的抗冻能力。结果表明,2种冷冻方法对小鼠GV期卵母细胞解冻后形态正常率和存活率无显著影响(P>0.05)。冷冻保护剂种类对小鼠GV期卵母细胞解冻后形态正常率无显著影响(P>0.05);但对存活率有显著影响,玻璃化冷冻采用二甲基亚砜作为冷冻保护液效果极显著优于甘油(P<0.01)。以冷冻效果较好的二甲基亚砜作为冷冻保护液,采用玻璃化冷冻不同发育阶段(GV期和MⅡ期)的小鼠卵母细胞,解冻后形态正常率无显著差异(P>0.05),但存活率GV期要显著优于MⅡ期卵母细胞(P<0.05)。 相似文献
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Tritrichomonas foetus is an extracellular parasite of the urogenital tract in cattle. It causes infertility and abortion, but there is no documented information on the susceptibility of bovine oocytes to the parasite, except by one article that claimed no effects of T. foetus on oocytes or embryos. The aim of the present study was to study the effects provoked by T. foetus when in interaction with bovine oocytes. Oocytes were obtained from cow ovaries and divided into two groups: (1) one group contained cumulus cells, whereas (2) a second group was denuded from these cells. Light microscopy, video microscopy and scanning electron microscopy (SEM) revealed that exposure of oocytes to T. foetus caused rapid adhesion of the trichomonads to cumulus cells and to the zona pellucida (ZP). Motile parasites were observed for 12 h. The ZP was completely damaged, and the parasites were able to infiltrate beneath the ZP and reached the oocytes directly when the oocytes were denuded of the cumulus cells. Both the oocytes and the cumulus cells exhibited morphological characteristics compatible with apoptosis after interaction with T. foetus, such as chromatin condensation, the presence of several cytoplasmic vacuoles, with intact cellular membranes and organelles. The results from this study demonstrate that when a large number of T. foetus interacts with oocytes in vitro damage and apoptosis are provoked in the cow's reproductive cells. The behavior of this parasite as one of the causes of cattle infertility is discussed. 相似文献
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家畜卵母细胞冷冻保存的研究进展 总被引：1，自引：1，他引：0
本文阐述了卵母细胞冷冻保存对体外受精、转基因动物研究及人类医疗方面的意义,论述了卵母细胞冷冻保存的发展过程、方法与机理及冷冻保护剂种类,通过揭示冷冻、解冻对细胞的作用,进一步探讨了各种冷冻保护剂及冻存方法对保持细胞生命力的功效。在当前卵母细胞冷冻研究中,玻璃化冷冻法发展迅速,并出现超快速玻璃化法。 相似文献
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Kim HJ Choi SH Son DS Cho SR Choe CY Kim YK Han MH Ryu IS Kim IC Kim IH Im KS Nagai T 《The Journal of reproduction and development》2006,52(5):633-638
This study was conducted to evaluate the effects of exposing porcine ovaries to 30-33 C during transportation for 4 h and subsequently room temperature (25 C) for 6 h of storage on in vitro maturation (IVM) and subsequent parthenogenetic development of oocytes collected from the ovaries. After IVM, oocytes having a tight oopalsm membrane and no signs of degeneration were exposed to Dulbecco's phosphate-buffered saline (DPBS) with 7% ethanol (v/v) for 7 min to induce parthenogenetic activation. Moreover, we also determined whether exposure of the collected oocytes to room temperature for 1, 2 and 4 h in DPBS or porcine follicular fluid (pFF) affected parthenogenetic development. When porcine ovaries were stored after transportation, oocytes collected from the stored ovaries showed a significantly higher rate of degeneration after 65 h of IVM (58.4%) and a significantly lower rate of cleavage after parthenogenetic activation (40.1%) than oocytes collected from ovaries immediately after transportation (38.9% and 47.4%, respectively). However, there was no significant difference in developmental rates to the morula and blastocyst stages between these two groups (14.4% and 14.3%, respectively). The duration of preservation, 1, 2, and 4 h, of oocytes in DPBS did not affect parthenogenetic development. In contrast, when preserved for 4 h in pFF, the developmental rates of the oocytes were significantly decreased. This suggested that some factor(s) in follicular fluid affects the developmental rate of oocytes with the passage of time in ambient conditions. These results suggest that even after 6 h storage of ovaries, oocytes having normal morphology after IVM have the same rate of parthenogenetic development as oocytes collected from ovaries just after 4 h of transportation, except for a lower cleavage rate, and that exposure of oocytes to room temperature for 4 h in DPBS does not affect their parthenogenetic developmental competence. 相似文献
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综述了哺乳动物卵母细胞孤雌生殖的主要研究进展,包括卵母细胞的成熟与激活、孤雌发育,以及游离Ca2 浓度的测定。为卵母细胞成熟体系、胚胎发育体系和胚胎干细胞体系的建立,以及体细胞核移植试验提供适宜的方法。 相似文献
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Ayano OI Hidetaka TASAKI Yasuhisa MUNAKATA Koumei SHIRASUNA Takehito KUWAYAMA Hisataka IWATA 《The Journal of reproduction and development》2015,61(3):191-197
In this study, we examined the effects of reconstructed oocyte–granulosa cell complexes (OGCs) on the development of porcine oocytes derived from early antral follicles (EAFs; 0.5–0.7 mm in diameter). When denuded oocytes were cocultured with granulosa cells derived from other EAFs, the oocytes and granulosa cells aggregated to form OGCs after 2 days of culture. After 14 days of culture, we compared cell number, oocyte diameter, and oocyte chromatin configuration in unmanipulated (natural) OGCs, reconstructed OGCs, and OGCs collected from antral follicles (AFs, 3.0–6.0 mm in diameter). The diameters of oocytes from reconstructed OGCs grown in vitro were not different from those of oocytes from natural OGCs, although they were significantly smaller than those of oocytes from antral follicle (AF) OGCs. Oocyte chromatin configuration did not differ among the 3 OGC groups, but the oocyte nuclear maturation rate was lower in the reconstructed OGCs and higher in
the AF OGCs. However, when the in vitro culture period for the reconstructed OGCs was extended by 2 days, the nuclear maturation rate of oocytes from reconstructed OGCs was similar to that of oocytes from natural OGCs. In addition, blastocysts were successfully obtained from oocytes from reconstructed OGCs. In conclusion, we established an innovative culture method that allows oocytes and granulosa cells from EAFs to reaggregate as reconstructed OGCs, which yield oocytes with the ability to develop to the blastocyst stage. 相似文献