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利用非手术法活体采卵技术结合体外受精技术和胚胎移植技术,能够更大限度的发挥良种母牛的繁殖潜力,并能够用于患有生殖道疾病良种母牛的扩繁,使良种资源得以充分利用;通过活体采卵技术还可以为核移植、转基因、胚胎干细胞移植等生物工程技术提供必要的材料。本文综述了年龄、发情周期、非手术法活体采卵方法等五个方面对牛非手术法活体采卵效果的影响,为探索更加有效的活体采卵程序和方法,提高非手术法活体采卵的生产效率提供依据。 相似文献
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为建立稳定高效的活体采卵-体外受精技术体系,提高体外胚胎生产效率,本研究先利用屠宰场采集的新鲜卵巢卵母细胞进行体外受精,通过胚胎发育潜力来筛选最佳的体外胚胎培养液;再进一步研究不同种公牛精液和供卵母牛对活体采卵-体外受精效率的影响。结果显示,CR1aa培养液和mCR1aa培养液卵裂率差异不显著(P>0.05),但mCR1aa组的囊胚发育率显著提高(28.1% vs 20.6%,P<0.05);选取的3头荷斯坦种公牛精液的活体采卵-体外受精胚胎的卵裂率差异不显著(P>0.05),但1号种公牛精液体外受精后囊胚率(38.7%)显著高于2号和3号(23.8%&22.9%)(P<0.05);随机选择的3头活体采卵供体母牛(H1、H2、H3)获得的头均可用卵母细胞数无显著差异,但H1和H2供体母牛体外受精胚胎的卵裂率和囊胚率均显著高于H3供体牛(P<0.05),且H1供体牛体外受精囊胚率显著高于H2供体牛(P<0.05)。结果表明,mCR1aa培养液能显著提高体外受精囊胚发育率,适用于体外胚胎生产;种公牛精液和供体母牛个体差异会直接影响活体采卵-体外受精胚胎的生产效率,为奶牛活体采卵-体外受精生产技术体系的优化提供参考。 相似文献
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Yindee M Techakumphu M Lohachit C Sirivaidyapong S Na-Chiangmai A Roelen BA Colenbrander B 《Reproduction in domestic animals》2011,46(5):824-831
This study was designed with the final goal of improving in vitro embryo production in the Thai swamp buffalo (Bubalus bubalis carabensis). Oocytes were collected by ovum pick-up (OPU) from six non-lactating multiparous swamp buffalo twice per week for 10 consecutive sessions followed by once-weekly collection for 10 consecutive sessions without hormone stimulation. In addition, oocytes were collected from slaughterhouse ovaries that were classified as follows: ovaries from non-pregnant cows with a visible corpus luteum (NPCL); pregnant cows with a corpus luteum (P); and non-pregnant cows without a corpus luteum (NP). Follicles in each group of ovaries were categorized as small (2-4 mm), medium-sized (5-8 mm) or large follicles (≥ 9 mm). The quality of the oocytes was assessed by their capacity to undergo in vitro maturation. The total number of observed follicles per session (all sizes combined) was larger in the once-weekly OPU group compared with the twice-weekly OPU group. In particular, the numbers of small and large follicles were higher in the once-weekly OPU group (5.2 ± 0.7 and 0.9 ± 0.2, respectively) than in the twice-weekly OPU group (3.9 ± 0.5 and 0.5 ± 0.1). The number of medium-sized follicles did not differ between the groups. The percentages of oocytes with an abnormal spindle morphology were not different between oocytes from the twice-weekly (30.0%) and the once-weekly (28.6%) OPU groups. A higher percentage of oocytes obtained in vitro (49.5%) exhibited nuclear abnormalities compared with those obtained in vivo (≤34.8%) after in vitro maturation. In conclusion, oocytes can be successfully collected by OPU in the swamp buffalo, without hormonal pretreatment, and per week more good-quality oocytes can be collected by twice-weekly OPU. In addition, oocytes collected from slaughterhouse ovaries can be used with the reproductive status of the cow having no influence on the maturation competence of oocytes. 相似文献
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Cytoskeletal and mitochondrial properties of bovine oocytes obtained by Ovum Pick‐Up: the effects of follicle stimulation and in vitro maturation 下载免费PDF全文
Tamás Somfai Satoko Matoba Yasushi Inaba Michiko Nakai Kei Imai Takashi Nagai Masaya Geshi 《Animal Science Journal》2015,86(12):970-980
Follicle stimulation by follicular stimulating hormone (FSH) is known to improve developmental competence of bovine oocytes obtained by Ovum Pick‐Up (OPU); however, the exact factors in oocytes affected by this treatment have remained unclear. We compared in vitro matured (IVM) oocytes obtained at the immature stage from cows by OPU either without or with stimulation with FSH (non‐stimulated and stimulated OPU, respectively) to those obtained by superstimulation and in vivo maturation in terms of cytoskeleton morphology, mitochondrial distribution, intracellular adenosine triphosphate (ATP) content and H2O2 levels at the metaphase‐II stage and intracellular Ca2+ levels after in vitro fertilization (IVF). Confocal microscopy after immunostaining revealed reduced size of the meiotic spindle, associated with increased tendencies of microfilament degradation and insufficient mitochondrial re‐distribution in non‐stimulated OPU‐derived IVM oocytes compared with those collected by stimulated OPU, which in turn resembled in vivo matured oocytes. However, there was no difference in mitochondrial functions between oocytes obtained by stimulated or non‐stimulated OPU in terms of ATP content, cytoplasmic H2O2 levels, base Ca2+ levels and the frequencies and amplitudes of Ca2+ oscillations after IVF. Larger size of metaphase spindles in oocytes obtained by stimulated OPU may reflect and potentially contribute to their high developmental competence. 相似文献
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Assisted reproduction technologies (ART) are well developed in humans and cattle and are gaining momentum also in the equine industry because of the fact that the mare does not respond to superovulation but can donate large numbers of oocytes through ovum pick up (OPU). After collection, the oocytes can be fertilized by intracytoplasmic sperm injection (ICSI) using a variety of stallion semen samples, even of poor quality, and the resulting embryos can establish high pregnancy rates after cryopreservation and transfer. The discoveries that equine oocytes can be held at room temperature without loss of viability and that an increase in vitro maturation time can double the number of embryos produced are fueling the uptake of the OPU technique by several clinics that are shipping oocytes of their client’s mares to specialized ICSI laboratories for embryo production and freezing. In this article, we present a retrospective analysis of 10 years of work at Avantea with a special focus on the last 3 years. Based on our data, an average production of 1.7 to 2 embryos per OPU-ICSI procedure can be obtained from warmblood donor mares with a pregnancy rate of 70% and a foaling rate in excess of 50%. OPU-ICSI offers the added value of freezing embryos that allows the development of embryo commercialization worldwide to the benefit of top horse breeders who are endorsing this technology as never before. 相似文献
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水牛分离精子与不同来源卵母细胞体外受精的效果研究 总被引:1,自引:1,他引:0
本研究的目的是探讨水牛分离精子与不同来源(活体采卵或屠宰场卵巢采卵)卵母细胞体外受精的效果。活体采卵是选用20头空怀河流型母水牛(其中摩拉母牛12头,尼里-拉菲母水牛8头)每间隔3 d采卵1次,连续采卵5~6周,活体采集卵母细胞;屠宰场卵巢采卵是收集屠宰场水牛卵巢,用10 mL注射器连接18 G针头吸取水牛卵巢上可视的卵泡来收集卵母细胞。将收集的AB级水牛卵母细胞在相同的条件下进行体外成熟、然后用分离或未分离精子进行体外受精以及体外培养至囊胚。结果发现:活体采卵组和屠宰场收集的水牛卵母细胞组用分离精子受精分裂率和囊胚率没有差异(P>0.05);分离精子和未分离精子的体外受精分裂率和囊胚率也没有差异(P>0.05)。由此说明,水牛分离精子可以用于体外生产性控胚胎。 相似文献
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The objective of this study was to investigate (i) the effect of two different ovum pick-up (OPU) schemes (once vs twice weekly aspirations) on oocyte recovery rate, quality and subsequent in vitro embryo development, (ii) the influence of days post-partum on oocyte recovery and (iii) possible differences in OPU results from two different herds. In group A, OPU was performed twice weekly in two Holstein Friesian (HF) and three Danish Red and White (DRW) cows from a private herd. In the research herd, two groups of eight HF cows were investigated: group B (OPU once weekly) and group C (OPU twice weekly). The collected oocytes were subsequently submitted to in vitro embryo production. More oocytes were recovered from the private herd when compared with the research herd. In the research herd, the twice weekly scheme aspirated more oocytes than the once weekly scheme. The quality of the retrieved oocytes was significantly different between groups B and C but not between groups A and C, and HF cows yielded higher quality oocytes than DRW cows (p = 0.029). Oocytes from group C showed higher level of embryonic development than group B oocytes. No differences in blastocyst rates were observed between groups A and C. Session affected the number of retrieved oocytes and subsequent developmental rates, with these being lower in the first compared with the last sessions. Finally, there was no significant effect of days post-partum in the number and quality of the retrieved oocytes, likely because of the small group size and high variation between sessions. 相似文献
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本研究比较了不同来源的水牛卵母细胞的体外受精及其胚胎发育。对10头摩拉母水牛连续6周进行活体采卵,共采集292枚卵母细胞,头均回收卵母细胞4.87枚,头均A级卵数为3.07枚,从屠宰场收集74头水牛卵巢共采集559枚卵母细胞,头均回收卵母细胞和A级卵母细胞分别为7.55枚和5.20枚,均显著高于活体采集的水牛卵母细胞(P<0.01)。将收集的AB级水牛卵母细胞在相同的条件下进行体外成熟、体外受精和体外培养至囊胚。结果表明,活体采卵组和屠宰水牛卵母细胞组受精分裂率差异不显著,分别为54.81%和57.73%。屠宰水牛卵母细胞组的囊胚率则高于活体采卵组(28.78%vs21.34%,P<0.05)。利用两组的胚胎进行常规法冷冻保存,冻胚解冻后的存活率差异不显著(P>0.05)。 相似文献
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Toshinori OIKAWA Tomoko ITAHASHI Takashi NUMABE 《The Journal of reproduction and development》2016,62(1):11-16
The purpose of this study was to determine whether dithiothreitol (DTT) treatment of sperm and ethanol
activation improve embryo production by intracytoplasmic sperm injection (ICSI). Further, we compared ICSI
with standard in vitro fertilization (IVF) in oocytes obtained from cattle. We demonstrated
that DTT reduced the disulfide bond in the bovine sperm head. Using oocytes obtained from a slaughterhouse,
ICSI-DTT treatment without ethanol showed the highest rate of blastocyst formation. We applied these results
to fertilization using ovum pick-up (OPU). Eleven Japanese black cattle served as donors for OPU plus standard
IVF (OPU-IVF). Of them, four donors with low embryo development rates were selected to determine whether
embryo development was enhanced by OPU plus ICSI (OPU-ICSI). We assessed effects on embryo development
following IVF and ICSI in oocytes obtained using OPU. Blastocyst rates were significantly higher for OPU-ICSI
than for OPU-IVF. Our results suggest that OPU-ICSI improves the blastocyst development rate in donors with
low embryo production compared with the standard OPU-IVF. 相似文献
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Hirata T Sato M Sasaki S Sasaki O Osawa T 《The Journal of reproduction and development》2008,54(5):346-351
We investigated whether suckling would affect embryo production of cows bred by timed artificial insemination (TAI) following an ovulation synchronization protocol combined with ovum pick-up and progesterone releasing intravaginal device (OPU-PRID-TAI protocol). The number of oocytes and transferable embryos collected by repeated OPU, performed before and after TAI, were recorded. A total of 14 Japanese Black cows were divided into weaned (n=7) and suckled groups (n=7). All 14 cows were treated with OPU on day 0 (the first day of treatment) and then with a PRID for 9 days. Prostaglandin F(2alpha) analog was administered on day 7, GnRH analog was administered on day 10 (36 h after removal of the PRID) and TAI was performed 12 h later. Ovulation was confirmed by palpation per rectum the following day. After TAI, additional OPU sessions were performed on days 18, 25 and 32. The synchronized ovulation rates of the weaned and suckled groups were 100 and 85.7%, and the conception rates were 71.4 and 42.9%, respectively. Immature oocytes were fertilized and cultured in vitro. The numbers of oocytes collected and blastocysts generated were similar between the individual OPU sessions in both groups. However, the total numbers of oocytes collected, cultured oocytes, cleavage embryos and blastocysts as well as the proportions of cleavage embryos and blastocysts to cultured oocytes were all significantly (P<0.05) greater in the weaned group compared with the suckled group. These results suggest that the OPU-PRID-TAI protocol has the potential to produce a significant number of good-quality embryos in vitro after repeated OPU in early postpartum weaned Japanese Black cows. To collect more oocytes and produce more embryos, we suggest that calves be removed from cows scheduled for treatment using this protocol. 相似文献