首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献，搜索用时 140 毫秒
1.
Over the last several decades, a number of therapies have been developed that manipulate ovarian follicle growth to improve oocyte quality and conception rates in cattle. Various strategies have been proposed to improve the responses to reproductive biotechnologies following timed artificial insemination (TAI), superovulation (SOV) or ovum pickup (OPU) programmes. During TAI protocols, final follicular growth and size of the ovulatory follicle are key factors that may significantly influence oocyte quality, ovulation, the uterine environment and consequently pregnancy outcomes. Progesterone concentrations during SOV protocols influence follicular growth, oocyte quality and embryo quality; therefore, several adjustments to SOV protocols have been proposed depending on the animal category and breed. In addition, the success of in vitro embryo production is directly related to the number and quality of cumulus oocyte complexes harvested by OPU. Control of follicle development has a significant impact on the OPU outcome. This article discusses a number of key points related to the manipulation of ovarian follicular growth to maximize oocyte quality and improve conception rates following TAI and embryo transfer of in vivo- and in vitro-derived embryos in cattle.  相似文献   

2.
In vivo ovum pick-up (OPU) in sheep may be improved with a proper choice of aspiration elements (needle and tubing) and aspiration vacuum pressure. In the present study, two experiments were carried out. In Expt 1, visible follicles in ovaries of slaughtered ewes (treated separately according to their diameters: small<3 mm, medium 3-5 mm and large>5 mm) were aspirated using different combinations of the three studied factors such as aspiration flow rate (10, 20, 30, 40 and 50 ml water/min), needle gauge (18 and 20 G) and tubing inner diameter (1, 2 or 3 mm internal diameter). In Expt 2, a study with two 18 G needles of different lengths (18 G: 82 mm; 18 GL: 600 mm) was carried out, using ovaries obtained post-mortem, and performing in vivo laparoscopic follicular aspiration on ewes. We considered good quality oocytes as those with both complete compact cumulus and a homogeneous cytoplasm. Recovery rate, proportion of good quality oocytes (good quality oocytes/100 oocytes recovered) and overall efficiency (good quality oocytes/100 follicles aspirated) were noted. In Expt 1, aspiration flow rate affect remarkable proportion of good quality oocytes (69.5%, 50.5%, 44.8%, 36.5% and 28.3% for flows from 10 to 50 ml/min respectively, p<0.05). Needle gauge did not affect aspiration device efficiency. Thin and intermediate tubings were more effective (overall efficiency rates: 34.9%, 32.3% and 28.1% for 1, 2 and 3 mm respectively, p<0.05). Follicle size did not affect recovery rate, but proportion of good quality oocytes was higher for large (77.9%) and medium (64.4%) follicles (p<0.05). Finally, some combinations of the aspiration device showed greater effectiveness. In Expt 2, needle length did not influence recovery rate, but good quality oocytes rate was significantly modified both post-mortem and in vivo (good quality rate for 18 G vs 18 GL needles: 69.5% vs 47.7% and 58.1% vs 25.4%, post-mortem and in vivo respectively, p<0.05). We conclude that low-aspiration flow rates (10 and 20 ml/min) with thin or intermediate tubings (1 and 2 mm), and any short needle (18 G or 20 G) are the most adequate aspiration factors for OPU in sheep.  相似文献   

3.
  相似文献   

4.
The aim of this investigation was to examine the chromatin configuration of the nucleus, pattern of mitochondrial aggregation and mitochondrial activity in parallel studies in the same horse oocytes. Horse oocytes recovered by ultrasound-guided follicle aspiration in vivo were classified according to two main initial cumulus morphologies as having compact or expanded cumulus. The percentage of oocytes with a diplotene meiotic configuration at the time of recovery from the follicles was highest in compact oocytes. Oocytes with expanded cumulus layers at the time of recovery matured more rapidly in vitro and reached a proportion >50% at the metaphase II stage (M 2) sooner during in vitro maturation (IVM), than did compact oocytes. The mitochondrial aggregation pattern changed from finely distributed (Type 1) through crystalline (Type 2) to an aggregated, granulated appearance (Type 3) during IVM. The pattern of mitochondrial aggregation at the time of recovery was associated with the initial cumulus morphology of the oocyte, in that compact oocytes had a higher proportion of Type 1 aggregation, whereas expanded oocytes had a higher proportion of Type 3. The fluorescence intensity of metabolic active mitochondria, measured by fluorescence intensity (Em 570) per oocyte after MitoTracker CMTM Ros orange labelling, increased in the oocytes during IVM and depended on initial cumulus investment. Oocytes with the granulated type of aggregated mitochondria Type 3 had the highest level of metabolic activity and were in more progressed stages of meiosis (A 1-M 2). Oocytes initially having expanded layers of cumulus reached significantly higher levels of mitochondrial activity after IVM than did oocytes initially having compact cumuli. During resumption of meiosis the mitochondrial activity of oocytes with initially expanded cumulus increased continuously up to M 2, whereas in oocytes from compact cumulus-oocyte complex (COC), the activity declined after A 1/T 1 stages of meiosis.  相似文献   

5.
The aim of this retrospective study was to compare the number of follicles, cumulus oocyte complexes (COCs) and cultured In Vitro Produced (IVP) embryos obtained from 1396 non‐stimulated Ovum Pick‐up (OPU) sessions on 81 donor animals in a twice weekly OPU scheme. Results were obtained from 640 sessions following FSH‐LH superstimulation, on 112 donors subjected to OPU once every 2 weeks. The stimulation protocol started with the insertion of an ear implant containing 3 mg norgestomet (Crestar, Intervet, Belgium) 8 days before puncture (day ?8). The dominant follicle was ablated by ultrasound‐guided follicle puncture on day ?6. On day ?3 and day ?2, cows were injected with FSH (Ovagen, ICP) twice daily (8 am to 8 pm ), i.e. a total dose of 160 μg FSH and 40 μg LG per donor per stimulation cycle. Animals were punctured 48 h after the last FSH injection (day 0). Progesterone implants were removed the next day. Stimulated donor cows were treated with this protocol at 14‐day intervals. Follicles were visualized with a Dynamic Imaging ultrasound scanner, equipped with a 6.5 MHz sectorial probe. Follicles were punctured with 55 cm long, 18 gauge needles at an aspiration pressure corresponding to a flow rate of 15 ml/min. Cumulus oocyte complexes were recovered and processed in a routine IVF set‐up. Results demonstrate that, expressed per session, FSH stimulation prior to OPU increases production efficiency with significantly more follicles punctured and oocytes retrieved. However, when overall results during comparable 2‐week periods are considered (four non‐stimulated sessions vs one stimulated), more follicles are punctured and more oocytes are retrieved using the non‐stimulated protocol. No significant differences in the number of cultured embryos could be detected, indicating that FSH/LH stimulation prior to OPU might have a positive effect on in vitro oocyte developmental competence as more embryos are cultured with less, presumably better‐quality, oocytes.  相似文献   

6.
This study examined the effect of treating mares with equine pituitary extract (EPE) in combination with human chorionic gonadotropin (hCG; EPE/hCG) on the recovery rate of immature oocytes by ovum pick-up (OPU) and on oocyte morphology. Ten mares were subjected to each of two treatments in a random sequence: superstimulated with EPE (25 mg, twice daily) and treated with hCG (2,500 IU) or control (no exogenous treatment). The cytoplasmic morphology of oocytes recovered was evaluated through transmission electron microscopy. Follicular fluid was collected at aspiration for progesterone analysis, which was performed by radioimmunoassay. The EPE/hCG did not increase the oocyte recovery rate from immature follicles when compared with the controls (15.5% and 16.7%, respectively). A significantly higher oocyte recovery rate per mare was observed (70% versus 50%). However, precocious granulosa cell expansion was observed with EPE/hCG treatment in contrast to the control (64.4% and 33% of follicles with expanded cumulus, respectively), and increased intrafollicular progesterone concentration was also seen (158.80 ng/mL versus 82.05 ng/mL). The ultrastructural analysis of oocytes from both groups showed morphologic features related to immaturity. Numerous vesicles containing cortical granules were found, distributed in clusters into the cytoplasm, and junctional complexes were still seen between oocyte and granulosa cells. In conclusion, EPE/hCG treatment induced some follicular modifications, but the recovery rate was not increased. All oocytes examined presented signs of immaturity.  相似文献   

7.
The objective of the experiment was to study oocyte recovery by transvaginal, ultrasound-guided, follicle aspiration, from Thai swamp buffalo using different vacuum pressures. Six adult buffalo heifers, aged 2.5-3.0 yrs were treated with a total dose of 280 mg FSH, given twice a day in a divided doses over a three day period (60/60 mg, 50/50 mg, 30/30 mg) at d7 after progesterone implant. Three vacuum pressures were used; 100 (n=12), 80 (n=12) and 60 mmHg (n=12) and all of the pressures were performed in each animal. The animals were treated repeatedly and collection took place using 2 sets of each pressure every 2 months, giving a total of 36 collections from each animal. The oocyte recovery rates from each pressure were 81.2% (69/85) 79.1% (53/67) and 90.3% (93/103) for 100, 80 and 60 mmHg respectively. The number of oocytes collected per donor were 5.33 +/- 3.27, 4.42 +/- 2.71 and 7.75 +/- 4.31 respectively. The quality of the oocytes did not improved with the lower vacuum pressure. In conclusion, the application of FSH pretreatment improves the yield of oocytes from Thai, swamp buffalo heifers after gonadotropin treatment when using the vacuum pressures between 60-100 mmHg.  相似文献   

8.
Transvaginal ultrasound-guided follicle aspiration is one method of obtaining recipient oocytes for equine somatic cell nuclear transfer (SCNT). This study was conducted: (1) to evaluate the possibility of oocyte aspiration from pre-ovulatory follicles using a short disposable needle system (14-G) by comparing the oocyte recovery rate with that of a long double lumen needle (12-G); (2) to investigate the developmental competence of recovered oocytes after SCNT and embryo transfer. The recovery rates with the short disposable needle vs. the long needle were not significantly different (47.5% and 35.0%, respectively). Twenty-six SCNT embryos were transferred to 13 mares, and one mare delivered a live offspring at Day 342. There was a perfect identity match between the cloned foal and the cell donor after analysis of microsatellite DNA, and the mitochondrial DNA of the cloned foal was identical with that of the oocyte donor. These results demonstrated that the short disposable needle system can be used to recover oocytes to use as cytoplasts for SCNT, in the production of cloned foals and for other applications in equine embryology  相似文献   

9.
The proportion of transferable beef embryos obtained after superovulation, follicle aspiration, and in vitro maturation and fertilization has been small. To seek possible explanations, cows on different planes of nutrition were treated with exogenous gonadotropin and oocytes were isolated from their ovaries. The record for each oocyte included characteristics of the follicle, ovary, and cow from which it was obtained and the response to in vitro maturation, fertilization, and development. The sample was used to obtain estimates of the relationships among the variables. The logistic function with the probability of normal development as the dependent variable was the basic equation of the statistical model. When an explanatory variable was itself a result of the biological system, an equation explaining variation therein was added to the model. Had equations representing endogenous regressors not been added to the model a simple, single equation would have represented oocyte development response; given an oocyte at aspiration only one variable, cumulus quantity, was found to condition the probability of normal development directly. However, the complete model included four additional equations: 1) the probability that an oocyte was recovered at aspiration was conditional on the plane of nutritional treatment and progesterone concentration in follicular fluid; 2) cumulus quantity was conditional on the presence on a corpus luteum, follicle size, and progesterone concentration; 3) progesterone concentration was dependent on plane of nutrition; and 4) corpus luteum was conditional on plane of nutrition. The estimated model provided some insight into the complexity of oocyte development response and the role nutrition may play.  相似文献   

10.
本试验利用B型超声波导引活体采集42头奶水牛的卵母细胞,每头牛每周采卵2次,共采卵96次,卵母细胞回收率67%,可用卵母细胞数平均为3.1,成熟培养的囊胚率平均为24.7%.对水牛的卵巢观察结果表明活采对水牛的健康及生殖机能不产生影响.  相似文献   

11.
Cattle induced to ovulate a small, physiologically immature preovulatory follicle had reduced oocyte developmental competence that resulted in decreased embryo cleavage and day 7 embryo quality compared with animals induced to ovulate a more advanced follicle. RNA-sequencing was performed on oocytes and their corresponding cumulus cells approximately 23 h after gonadotropin-releasing hormone (GnRH) administration to induce the preovulatory gonadotropin surge suggested reduced capacity for glucose metabolism and oxidative phosphorylation in the cumulus cells and oocytes from follicles ≤11.7 mm, respectively. We hypothesized that induced ovulation of a small, physiologically immature preovulatory follicle results in a suboptimal follicular microenvironment and reduced oocyte metabolic capacity. We performed a study with the objective to determine the impact of preovulatory follicle diameter and serum estradiol concentration at GnRH administration on oocyte metabolic competence and follicular fluid metabolome profiles. We synchronized the development of a preovulatory follicle and collected the follicle contents via transvaginal aspiration approximately 19 h after GnRH administration in lactating beef cows (n = 319). We determined ATP levels and mitochondrial DNA (mtDNA) copy number in 110 oocytes and performed ultra-high-performance liquid chromatography–high resolution mass spectrometry metabolomic studies on 45 follicular fluid samples. Intraoocyte ATP and the amount of ATP produced per mtDNA copy number were associated with serum estradiol concentration at GnRH and time from GnRH administration to follicle aspiration (P < 0.05). mtDNA copy number was not related to follicle diameter at GnRH, serum estradiol concentration at GnRH, or any potential covariates (P > 0.10). We detected 90 metabolites in the aspirated follicular fluid. We identified 22 metabolites associated with serum estradiol concentration at GnRH and 63 metabolites associated with follicular fluid progesterone concentration at the time of follicle aspiration (FDR < 0.10). Pathway enrichment analysis of significant metabolites suggested altered proteinogenesis, citric acid cycle, and pyrimidine metabolism in follicles of reduced estrogenic capacity pre-gonadotropin surge or reduced progesterone production by the time of follicle aspiration.  相似文献   

12.
During oocyte growth and follicle development, oocytes closely communicate with cumulus cells. We examined the effects of oocyte-derived growth factors, growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), on the growth and acquisition of meiotic competence of porcine oocytes collected from early antral follicles (1.2–1.5 mm). First, we confirmed that GDF9 and BMP15 mRNAs were expressed almost exclusively in the oocytes. Oocyte–cumulus cell complexes (OCCs) collected from early antral follicles were cultured in growth medium supplemented with 0–100 ng/ml of GDF9 or BMP15 for 5 days. GDF9 dose-dependently increased the OCC diameter, while BMP15 did not. GDF9 and BMP15 had no significant effects on oocyte growth (P > 0.05). When OCCs that had been cultured with 50 and 100 ng/ml BMP15 were subjected to a subsequent maturation culture, they expanded fully by gonadotropic stimulation and 49% and 61% of oocytes matured to metaphase II (MII), respectively. In contrast, GDF9 did not promote cumulus expansion, and < 10% of oocytes matured to MII. Based on the difference in cumulus expansion, we compared the expression of luteinizing hormone/choriogonadotropin receptor (LHCGR) and follicle stimulating hormone receptor (FSHR) mRNAs in cumulus cells. The level of LHCGR mRNA was increased in cumulus cells of the BMP15 group, although there were no significant differences in FSHR mRNA levels among the groups. These results suggest that GDF9 promotes the growth of OCCs and that BMP15 promotes LHCGR mRNA expression in cumulus cells during oocyte growth culture, which may contribute to cumulus expansion and oocyte maturation.  相似文献   

13.
Co‐culture of cumulus‐oocyte complexes (COCs) with denuded oocytes (DOs) during in vitro maturation (IVM) was reported to improve the developmental competence of oocytes via oocyte‐secreted factors in cattle. The aim of the present study was to investigate if addition of DOs during IVM can improve in vitro fertilization (IVF) and in vitro culture (IVC) results for oocytes in a defined in vitro production system in pigs. The maturation medium was porcine oocyte medium supplemented with gonadotropins, dbcAMP and β‐mercaptoethanol. Cumulus‐oocyte complexes were matured without DOs or with DOs in different ratios (9 COC, 9 COC+16 DO and 9 COC+36 DO). Consequently; oocytes were subjected to IVF as intact COCs or after denudation to examine if DO addition during IVM would affect cumulus or oocyte properties. After fertilization, penetration and normal fertilization rates of zygotes were not different between all tested groups irrespective of denudation before IVF. When zygotes were cultured for 6 days, no difference could be observed between all treatment groups in cleavage rate, blastocyst rate and cell number per blastocyst. In conclusion, irrespective of the ratio, co‐culture with DOs during IVM did not improve fertilization parameters and embryo development of cumulus‐enclosed porcine oocytes in a defined system.  相似文献   

14.
  相似文献   

15.
  相似文献   

16.
Contents: Bovine follicular oocytes were matured and fertilize din vitro. The frequency of penetration and subsequent embryonic development were improved considerably, for oocytes cultured in larger volumes allowing larger oocyte groups as compared to the culture of 2 oocytes within 30 μl drops. The effects of follicle stimulating hormone (FSH) and luteinizing hormone (LH), present during in vitro maturation, were studied in terms of cumulus expansion, oocyte penetration, male pronucleus formation and embryonic development. Cumulus expansion including mucification was induced by both hormones. Scanning electron microgfaphs revealed that storage of LH as a frozen solution over a long time period (10 months), destroyed its ability to stimulate cumulus mucification, whereas uncoupling of the cumulus cell processes still occurred. LH caused an increase in the percentage of penetrated oocytes with incomplete sperm head decondensation. This effect was also lost after long term storage. Teh resulting total penetration frequency as well as the proportion of oocytes with both pronuclei formed was now similar to that observed with oocytes matured with fresh LH or FSH. Embryonic development was not altered by the replacement of FSH by LH during in vitro maturation .  相似文献   

17.
The aim of this study was to determine the optimal maturation culture period of ovum pick up (OPU)‐derived cumulus oocytes complexes (COCs) in relation to their developmental capacity. Embryo production, embryo cryotolerance, post‐transfer embryonic survival and calf characteristics such as gestation length, birthweight and sex ratio were investigated. This retrospective study covers the analyses of ovum pick up –in vitro production and calving results from a commercial programme that took place between March 1994 and September 2004. Donors were both heifers (of which approximately 90% pregnant) and cows (of which approximately 10% pregnant). Embryo production analyses were based on 7800 OPU sessions conducted from January 1995 until January 1999. Analyses of calving rate were based on 13 468 embryo transfers performed during January 1995 until May 2002. Analyses on calf characteristics were based on 2162 calves born between March 1994 and September 2004. The in vitro maturation culture period ranged from 16 to 28 h. The mean production rate of transferable embryos was 16.5% (1.2 embryos per OPU session). Length of maturation culture period did not affect the production of transferable embryos. Mean calving rate was 40.9% and 38.7% for fresh and frozen/thawed embryos, respectively. Calving rate was not affected by the maturation culture period. Mean birthweight, gestation length and proportion of male calves were 46 kg, 281.9 days and 52.8%, respectively. Maturation culture period did not affect these variables. In conclusion, this study shows that the in vitro maturation culture period within the range of 16–28 h does not affect in vitro embryo production, embryo cryotolerance, post‐transfer embryonic survival and calf characteristics, suggesting that all COC batches collected by OPU on the same day, can be fertilized in one IVF session without a significant loss in the production from oocyte to calf.  相似文献   

18.
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   

19.
The present study was conducted to investigate the effects of attachment of cumulus cells to porcine oocytes during the process of maturation and fertilization on the nuclear maturation, fertilization and subsequent development after in vitro fertilization (IVF). In the first experiment, the cumulus cells were removed from cumulus-oocyte complexes (COCs) at 0, 24 and 42 h after the onset of maturation culture and were then cultured until reaching 42 h of cultivation. In the second experiment, COCs were denuded as described in the first experiment, then fertilized and cultured for 7 days. As a control, cumulus cells were allowed to maintain attachment to the oocytes until the end of IVF. The proportion of oocytes reaching metaphase II significantly increased with the delay in the removal treatment of cumulus cells. The proportion of normal fertilization gradually increased with delay in the removal treatment of cumulus cells from COCs until the end of IVF. However, no significant difference in the proportion of normal fertilization was found between the 42-h and control groups. The removal treatment of cumulus cells in the 0- and 24-h group significantly (p < 0.05) decreased the proportion of cleaved embryos when compared with the control, and none of them developed to the blastocyst stage. The proportion of development to the blastocyst stage was significantly higher (p < 0.05) in the control group than in the 42-h group (18.1% vs 12.4%; p < 0.05). The present study indicates that the attachment of cumulus cells to the oocyte during maturation and fertilization is important to support oocyte nuclear maturation, fertilization and subsequent embryo development. Particularly, the attachment of cumulus cells to the oocyte during IVF promotes embryonic development.  相似文献   

20.
This study aimed to evaluate gonadotropin secretion and the developmental competence of follicular oocytes in dairy cattle during the early postpartum (PP) period. The number of follicles developed after transvaginal ultrasound-guided ovum pick-up (OPU) and fertilizability of retrieved oocytes were compared between cows in which the first dominant follicle (DF) ovulated (ovulated group, n=4) and did not ovulate (non-ovulated group, n=3), and between early PP (early PP group, n=2) and after the resumption of the estrous cycle (cyclic group, n=2). Follicular ablation was performed 2-4 days after the detection of DF in the second follicular wave PP. OPU was repeated 3-5 times at 3 or 4-day intervals from 3-4 days after the follicular ablation. At OPU, the follicles were enumerated and all those > or = 5 mm in diameter were aspirated. Recovered oocytes were subjected to in vitro maturation and fertilization. Both criteria were similar between ovulated and non-ovulated groups, and between early PP and cyclic groups. These results suggest that FSH/LH secretions required for follicle recruitment and subsequent follicular growth during the early PP period are similar to those after resumption of the estrous cycle. They also indicate that follicular oocytes during the early PP period have developmental competence.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号