176 Effect of epigallocatechin-3-gallate on bovine oocyte in vitro maturation, fertilization, and development

2019 ◽  
Vol 31 (1) ◽  
pp. 212
Author(s):  
Y. Honkawa ◽  
Y. Gen ◽  
S.-H. Hyon ◽  
C. Kubota
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Polar Body ◽  
Sperm Concentration ◽  
P Value ◽  
Bovine Oocyte ◽  
Vitro Fertilization ◽  
Significant Difference ◽  
Maturation Rate

Epigallocatechin-3-gallate (EGCG) is a major ingredient of catechin polyphenols, and a strong antioxidant compound. Huang et al. (2018 Asian-australas. J. Anim. Sci.) reported that adding 50μM EGCG can improve the bovine oocyte maturation rate. In this research, we investigated the effect of EGCG supplementation on different periods in bovine IVF. Cumulus-oocyte complex (COC) collected from ovaries of slaughtered cows were cultured in maturation medium (20 to 30 oocytes per 100-µL droplet), which consisted of TCM-199 with Earle’s salts and 25mM HEPES supplemented with 10% (vol/vol) fetal bovine serum (FBS), 1µg mL−1 oestradiol, 0.02mg mL−1 FSH, and antibiotics at 38.5°C in a humidified atmosphere of 5% CO2 in air for 24h (in vitro maturation, IVM). After IVM, COC were fertilized in the fertilization medium (modified Brackett-Oliphant media supplemented with 10 µgmL−1 heparin, 10mM caffeine, and 3mg mL−1 BSA) for 6h using semen of one bull at final sperm concentration of 1×107 mL−1 (IVF). After IVF, COC were denuded and cultured in culture medium [CR1aa supplemented with 10% (vol/vol) FBS and antibiotics] at 38.5°C in a humidified atmosphere of 5% O2, 5% CO2, and 90%N2 for 8 days (in vitro culture, IVC). The EGCG was supplemented at 10, 25, 50, and 100M in IVM medium; 25 and 50 µM in IVF medium; and 50 and 100 µM in IVC medium. After 24h in IVM medium, COC were denuded by pipetting, fixed in 3:1 ethanol:acetic acid for 24h and then checked for nuclear and polar body by using aceto-orcein stain. After 18h in IVF, the pronucleus in zygote was fixed in 3:1 ethanol:acetic acid for 24h and checked by aceto-orcein staining. Embryo development was evaluated by counting the total number of embryos that had reached compacted morula by 6 to 8 days after IVF. Significant differences were analysed by chi-squared test and residual analysis. A P-value<0.05 was considered statistically significant. When EGCG was added to IVM, there was no significant difference of oocyte maturation rate between all concentrations (0v. 10v. 25v. 50v. 100 μM: 73.9% v. 56.7% v. 76.7% v. 72.7% v. 63.5%). When EGCG was added to IVF, there was no significant difference of fertilized rate (0v. 25v. 50 μM: 59.4% v. 73.7% v. 64.9%). When EGCG was added to IVC, there was no significant difference in development rate (0v. 50v. 100 μM: 26.2% v. 15.7% v. 22.0%). In this research, EGCG addition did not affect bovine in vitro fertilization.

scholarly journals Effect of follicle-stimulating hormone on Bligon goat oocyte maturation and embryonic development post in vitro fertilization

2020 ◽  
Vol 13 (11) ◽  
pp. 2443-2446
Author(s):  
Diah Tri Widayati ◽  
Mulyoto Pangestu
Keyword(s):  
In Vitro Fertilization ◽  
Embryonic Development ◽  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Follicle Stimulating Hormone ◽  
Fertilization In Vitro ◽  
Vitro Fertilization ◽  
Maturation Rate ◽  
Stimulating Hormone

Background and Aim: Bligon goat is a crossbreed between Etawah and Kacang goat. This crossbreed goat is mostly reared by small farmers. In vitro maturation allows female goat (does) contributes toward reproduction despite the fact that the animal has been slaughtered. The aim of this study was to determine the in vitro maturation rate of Bligon goat oocytes supplemented with follicle-stimulating hormone (FSH), and their ability for further embryonic development after in vitro fertilization. Materials and Methods: Experiment was conducted at the Laboratory of Animal Physiology and Reproduction, Faculty of Animal Science, Universitas Gadjah Mada, Yogyakarta, using Bligon goat ovaries obtained from local slaughterhouse around Yogyakarta. One thousand five hundred cumulus-oocyte complexes were matured for 24 h in tissue culture medium 199 supplemented with 50 IU/L FSH or without FSH (control). First, matured oocytes were evaluated its morphology based on the expansion of cumulus cells and PB1 extrusion. Next, 600 oocytes were then stained with 1% aceto-orcein to examine maturation based on changes in the configuration of chromosomes and nuclear membrane breakdown. Oocytes were considered mature when they reached metaphase II. To prove the ability of mature oocytes to develop into embryos, 900 oocytes were processed for fertilization in vitro. The data were analyzed using analysis of variance. Results: The results indicated that FSH supplementation significantly increased oocyte maturation rate (65.21±7.26 vs. 43.25±6.23%) as indicated by extrusion of PB1 and homologous chromosome pairing and lined in the equator. The rate of degeneration was lower in the FSH-supplemented medium (3.21±0.25 vs. 10.17±3.15%). The blastocyst stage of oocyte developed embryos was reached by 12.43±2.15% and 22.28±4.86% of the control and treatment groups, respectively. Conclusion: FSH supplementation significantly improves oocyte maturation and yields mature oocytes for future embryo development in vitro.

scholarly journals Use of Melatonin in the In Vitro Production of Bovine Embryos

2020 ◽  
Vol 21 ◽  
Author(s):  
Alan da Silva LIRA ◽  
Ricardo de Macedo CHAVES ◽  
Felipe de Jesus MORAES JUNIOR ◽  
Sergio Henrique COSTA JUNIOR ◽  
Brenda Karine Lima do AMARAL ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Control Group ◽  
Bovine Embryos ◽  
In Vitro Production ◽  
Maturation Medium ◽  
Significant Difference ◽  
Maturation Rate ◽  
Vitro Production

ABSTRACT We aimed to assess the effects of melatonin in the in vitro production of bovine embryos. Our experiment was conducted at the Laboratório de Reprodução Animal of the Universidade Estadual do Maranhão. The cumulus-oocyte complexes (COCs) were distributed among treatments at concentrations of 0, 10-1, 10-3 and 10-5 µMol/L melatonin. Our experiment was further divided into two: the first was to assess the effect of different concentrations of melatonin (treatments) on the maturation rate of COCs, and the second was to assess the effects of melatonin treatments on the in vitro production of bovine embryos. The results from the first experiment demonstrated no significant difference between the in vitro maturation rate of the cultivated COCs in treatments with melatonin. In the second experiment, however, melatonin treatments yielded statistically higher cleavage, morula and blastocyst rates in the 10-5 µM group (52.9%, 52.9%, and 35.3%, respectively), and lower rates in the 10-1 µM group (19.5%, 19.5% and 7.8%, respectively), compared to the others. The control group (no melatonin) and the 10-3 µM group showed similar results. We concluded that supplementation of melatonin in the in vitro maturation medium resulted in no improvement in the oocyte maturation rate, but in the in vitro production of embryos at different concentrations, the 10-5 µM group displayed better results, but with no improvement in the variables (P < 0.05).

336 SHEEP OOCYTES MATURED IN A SIMPLEX MEDIUM (HECM-1), AN ANSWER TO IN VITRO FERTILIZATION

2006 ◽  
Vol 18 (2) ◽  
pp. 275
Author(s):  
C. Navarro-Maldonado ◽  
Y. Ducolomb-Ramirez ◽  
A. Galindo-Rodriguez ◽  
A. Rosado-Garcia
Keyword(s):  
In Vitro Fertilization ◽  
Embryo Development ◽  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Culture Media ◽  
Sperm Quality ◽  
Complete Control ◽  
Vitro Fertilization ◽  
Significant Difference

In vitro maturation and in vitro fertilization (IVM and IVF) of mammalian oocytes still show unsatisfactory results when applied to the study of embryo development. This is probably due to inadequate information about the use of media components and supplements for oocyte maturation and to a discrepancy between results obtained by focusing strictly on oocyte maturation and those that are focused on IVF. A conventional medium that provides adequate results in studies of oocyte maturation (TCM-199) contains hypoxanthine, phosphate ions, and glucose, all known to inhibit embryo development in vitro in some species. In contrast, it has been shown that a simpler medium (HECM-9) increases embryo development in bovine although its use for oocyte maturation has not been defined. This medium contains taurine (an amino acid that reduces intracellular peroxide content) and is supplemented with polyvinyl alcohol (PVA) instead of using protein components, making it a simple defined medium that reduces variability in embryo development. Adding sodium panthothenate to media also confers cell protection against reactive oxygen species. Finally, supplements such as epidermal growth factor (EGF) increase the number of oocytes that complete maturation (Metaphase II, MII) and facilitate embryo development. An adequate combination of our knowledge about in vitro maturation and fertilization of oocytes, together with the requirements for embryo development, is important for the preparation of culture media to study regulatory mechanisms for embryo development and to increase the number of viable and normal term individuals. In this study we compared the effects of HECM-9 (containing panthothenate) vs. TCM-199 (both media supplemented with PVA, EGF, and FSH/LH) on the integrated processes involving IVM and IVF. No significant differences were found between the results obtained with these media in relation to oocyte maturation (65% MII for HECM-9 vs. 71% for TCM-199); however, those oocytes matured in HECM-9 showed a highly significant difference in in vitro fertilization using a conventional IVF medium (SOFm) (25% in HECM-9 vs. 6% in TCM-199). Maturation results were relatively low but in accordance with those reported by other groups, whereas IVF results are lower than those reported in the literature, perhaps because we have been using frozen and thawed samples and do not have complete control over the sperm quality. At present, we are extending our investigation using fresh semen samples.

155 Oocyte Maturation in Lyophilized In Vitro Maturation Medium as a Method to Increase the Medium’s Shelf-Life

2018 ◽  
Vol 30 (1) ◽  
pp. 217
Author(s):  
M. Rubessa ◽  
D. Weisgerber ◽  
S. Bessler ◽  
J. Bertels ◽  
B. Harley ◽  
...  
Keyword(s):  
Shelf Life ◽  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Polar Body ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Freeze Dried ◽  
Maturation Rate ◽  
The Impact

The in vitro production of bovine embryos has dramatically increased in recent years, and with it the demand of stable media with a long shelf-life. In this experiment we evaluated the impact of the freeze-dried in vitro maturation (IVM) medium (Mdry) on in vitro oocyte maturation. We compared the standard IVM and the Mdry media. Medium M199 was used as base for the IVM medium. The percentage of metaphase II oocytes and embryo production were evaluated. Media solutions (10 mL) were aliquoted into 50-mL conical tubes and lyophilized to form a powder concentrate using a Genesis freeze-dryer (VirTis, Gardener, NY, USA). Lyophilization consisted of a constant cooling from 20°C to –10°C at a constant rate of 1°C/min with a 2-h hold at –10°C before sublimation at 0°C. The Mdry medium was held at –80°C for 4 months (only serum and hormones were added before the incubation). When the IVM medium was rehydrated, the pH were adjusted to 7.4. The percentage of mature oocytes was evaluated after 24 h of maturation. The oocytes were stained with Hoechst 33342, and only oocytes with metaphase and a polar body were evaluated as matured. Abattoir-derived Holstein oocytes (n = 540) were in vitro matured (25–30/well in 400 µL) and fertilized with sexed semen, according to standard procedures (Rubessa et al. 2011 Theriogenology 76, 1347-1355). The oocytes were split for analysis (432 were used for IVP and 108 for maturation rate) over 6 replicates. Twenty hours after IVF, presumptive zygotes were cultured in SOF medium at 39°C with 5% CO2, 7% O2, and 88% N2. On Day 7, embryo yields were assessed. All recorded parameters were subjected to a Student’s t-test. The parameters compared were maturation rate, cleavage rate, blastocyst rate and the percentage of embryos cleaved. The α level was set at 0.05. All data were expressed as quadratic means and mean standard deviations. The results showed no differences between the 2 groups (75.9% v. 74.1%) (t = 0.37; SD = 12.69; P = 0.36; df = 5) when we compared the nuclear maturation; however, when we evaluated embryo production, we found the Mdry treatment had a higher cleavage percentage (t = 2.39; SD = 14.81; P = 0.02; df = 5) and total embryos produced (t = 2.49; SD = 5.6; P = 0.02; df = 5) compared with the control (Table 1.). These results showed that lyophilization can be a valid method to increase the shelf life of IVP media. More replicates must be done in order to understand why the freeze-dried media produced more embryos. Table 1.Mean (SD in parentheses) percentage cleavage and blastocysts

187 9-cis RETINOIC ACID IMPROVES MATURATION RATE AND ALTERS GENE EXPRESSION OF IN VITRO-MATURED OOCYTES IN EGYPTIAN BUFFALO

2017 ◽  
Vol 29 (1) ◽  
pp. 202
Author(s):  
A. Gad ◽  
S. Abu Hamed ◽  
M. Khalifa ◽  
A. El-Sayed ◽  
S. A. Swiefy ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Gene Expression ◽  
Retinoic Acid ◽  
Vitamin A ◽  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Polar Body ◽  
Control Group ◽  
Maturation Rate

Retinoic acid, a metabolite of vitamin A, regulates oocyte maturation through multiple mechanisms, including gene expression modulation or preventing oxidative stress. Effects of retinoic acid during oocyte maturation have been reported in several species; however, there have been no studies illustrating these effects in buffalo. Therefore, the objective of this study was to investigate the influence of 9-cis retinoic acid (9-cisRA), an active metabolite of vitamin A, on maturation rate and gene expression during in vitro maturation of buffalo oocytes. Cumulus-oocyte complexes (n = 360) were aspirated from surface follicles of Buffalo ovaries collected from local abattoirs and transported to the laboratory in physiological saline (0.9% NaCl) containing antibiotics (100 µg mL−1 of streptomycin sulfate and 100 IU mL−1 of penicillin) and maintained at 30°C. Grade A cumulus-oocyte complexes (evenly granulated cytoplasm and surrounded by multiple layers of cumulus cells) were randomly divided into 4 groups (90 oocytes/group) and allocated in TCM-199 medium supplemented with 10% fetal bovine serum, 0.2 mM sodium pyruvate, 50 μg mL−1 of gentamycin, and 10 μg mL−1 of FSH and contained 0 (control), 5, 50, or 200 nM of 9-cisRA for maturation. After 24 h, maturation rate was calculated as a percentage based on polar body extrusion. In addition, gene expression patterns were analysed for antioxidant related genes (SOD1, CAT, GPX4, HOMX1, and PRDX1) and oocyte quality-related genes (GDF9 and BMP15) using quantitative real-time PCR with GAPDH as a housekeeping gene. Fold changes (FC) were calculated using ΔΔCt method (FC ≥2; P < 0.05). The results showed that maturation rate (based on the extrusion of polar body) was significantly higher in 5 nM 9-cisRA oocyte group (49.4 ± 2.1%) compared with the control group (35 ± 1.8%); in contrast, the 200 nM 9-cisRA oocyte group showed the lowest maturation rate (27.2 ± 2.7%). However, the 50 nM 9-cisRA oocyte group showed no significant differences (31.2 ± 3.8%) compared with control group .Oocytes treated with 5 and 50 nM 9-cisRA during in vitro maturation showed significant up-regulation of SOD1 (3.4 and 3.08 FC), CAT (2.7 and 1.8 FC), and HOMX1 (4.5 and 4 FC), and significant down-regulation of BMP15 (−3.7 and −3.6 FC), respectively, compared with the control group. Moreover, GPX4, PRDX1, and GDF9 genes were highly expressed in the 50 nM compared with the control group (13.2, 10.4, and 1.8 FC, respectively). In contrast, the 200 nM 9-cisRA group showed significant down-regulation of CAT (−60.3 FC), GDF9 (−2.5 FC), and BMP15 (−9.7 FC) compared with the control group. In conclusion, these results suggested that a low concentration of 9-cisRA (5 nM) in maturation media can improves maturation rate of buffalo oocytes and up-regulates the expression of oxidative stress response-related genes.

165 In vitro maturation of ovine and caprine oocytes during breeding and nonbreeding seasons

2019 ◽  
Vol 31 (1) ◽  
pp. 207
Author(s):  
M. Markle ◽  
C. K. Mak ◽  
V. Medina ◽  
C. R. F. Pinto
Keyword(s):  
Breeding Season ◽  
In Vitro Maturation ◽  
Statistical Significance ◽  
Polar Body ◽  
Cumulus Cells ◽  
Nonbreeding Season ◽  
Nuclear Maturation ◽  
Significant Difference ◽  
Maturation Rate

The current study investigated the in vitro meiotic competence of ovine and caprine oocytes that underwent nuclear maturation during the breeding and nonbreeding seasons. We hypothesised that maturation rates of ovine and caprine oocyte would be significantly lower during the nonbreeding season. Ovine (Katahdin crossbred) and caprine (mainly Spanish crossbred) ovaries were collected from a local abattoir in the southern United States. Age of the animals was not determined. Cumulus-oocyte complexes (COC) were harvested by slicing the ovaries and searching using a stereomicroscope. Oocytes with more than 3 layers of unexpanded cumulus cells and with evenly granulated cytoplasm were selected for in vitro maturation (IVM). A commercial bovine IVM media (IVF Bioscience, Falmouth, United Kingdom) was used throughout the study. After 24h of IVM, ovine and caprine oocytes were denuded and oocytes with an extruded polar body (meiotic metaphase II oocytes) were considered to have reached nuclear maturation. The seasons in this study were defined as follows: breeding season=September to April and nonbreeding season=May to July. The presence of corpus hemorrhagicum or corpus luteum in at least 70% of the ovaries indicated the breeding season for the animals. Proportions of oocytes undergoing nuclear maturation were analysed using a two-tailed Chi-squared test. Statistical significance was set at P ≤ 0.05. The ovine maturation rate was 59% (65/111) and 49% (254/519) and the caprine maturation rate was 70% (39/56) and 40% (64/162) during the breeding and nonbreeding seasons, respectively. These results show a significant difference in nuclear maturation for caprine oocytes (P&lt;0.001) during the breeding and nonbreeding seasons; however, there was no significant difference in nuclear maturation for ovine oocytes (P=0.06) during the breeding and nonbreeding seasons. High environmental temperatures during the nonbreeding season may have had detrimental effects on oocyte nuclear maturation in caprine but not in ovine oocytes. Why oocytes from these 2 species differ on how they are adversely affected by season remains to be elucidated.

O-231 In vitro maturation of human immature (GV) oocytes after controlled ovarian hormonal stimulation with recombinant AMH in the maturation medium

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
J Bedenk ◽  
N Jančar ◽  
E Vrtačnik-Bokal ◽  
I Virant-Klun
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Polar Body ◽  
Time Lapse ◽  
Human Oocyte ◽  
Hormonal Stimulation ◽  
Maturation In Vitro ◽  
Maturation Medium ◽  
Maturation Rate

Abstract Study question Does the addition of recombinant AMH to the in vitro maturation (IVM) medium improve the maturation of GV oocytes after controlled ovarian hormonal stimulation? Summary answer Our results show that the addition of recombinant AMH to the in vitro maturation medium improves the maturation rate of GV oocytes. What is known already Anti-Müllerian hormone (AMH) is an important hormone involved in the process of sex differentiation during embryonic development. At the transition to the 21. century, more and more researchers have studied the role of AMH in ovarian function, especially its impact on folliculogenesis. AMH is becoming one of the main biomarkers of ovarian reserve and ovarian-specific disease, however, little is known about its effect on human oocyte maturation. Therefore, we matured immature GV (germinal vesicle) oocytes in IVM medium with recombinant AMH to assess its effect compared to the conventional IVM procedure with FSH and hCG. Study design, size, duration In this two-year prospective study, we compared the maturation rate of four groups of immature (GV) oocytes matured in maturation medium with added i) AMH (n = 15), ii) AMH+FSH+hCG (n = 44), iii) FSH+hCG (conventional; n = 22), and iv) hormone-free maturation medium (control; n = 15). Each oocyte was matured in vitro for a maximum of 28 hours and monitored by time-lapse microscopy to assess the time of GV breakdown (MI) and extrusion of the polar body (MII). Participants/materials, setting, methods Ninety-six GV oocytes of 46 patients (aged &lt; 38 years, involved in the ICSI programme) after short antagonist protocol of controlled ovarian hormonal stimulation were included after written informed consent. IVM of oocytes was performed in the MediCult IVM System (LAG and IVM medium, Cooper Surgical, Denmark) with added hormones, and in a CO2 incubator equipped with the PrimoVision time-lapse microscope (Vitrolife, Sweden). Main results and the role of chance IVM medium with added recombinant AMH gave the best result with all (100 %) oocytes matured in vitro. In conventional IVM medium with FSH and hCG, the oocyte maturation rate was poorer, with 68 % of oocytes matured in vitro. An even lower oocyte maturation rate (34 %) was observed in IVM medium with AMH, FSH and HCG, which might be explained by the antagonistic action of these hormones. In a group of control oocytes, 25 % of oocytes matured in vitro. The mean time to GV breakdown (MI stage) was 3.7 hours and to polar body release (MII stage) 20,5 hours. The time to MI stage was quite comparable in all groups of oocytes (3.5, 3.8 and 3.7 hours). There was a tendency for the polar body to be released later if AMH was added to the maturation medium (21.5 and 20.2 vs. 19.9 hours) but differences were not statistically significant, as revealed by Student’s t-test. In the control group of oocytes, these times were prolonged (4.2 and 22.2 hours) due to slow spontaneous maturation. These preliminary results demonstrate that AMH could directly affect the oocyte maturation in vitro. Limitations, reasons for caution The limitation is the relatively small number of oocytes included; GV oocytes accounted for less than 10 % of all oocytes in the in vitro fertilisation (ICSI) programme. Moreover, the proportion of GV oocytes spontaneously matured to MI stage before the start of the experiment and were therefore not included. Wider implications of the findings Based on our data, we believe that AMH directly affects human oocyte maturation in vitro. Despite the common knowledge that AMH regulates the recruitment of growing ovarian follicles, it appears that the addition of AMH to the maturation medium can improve the human oocyte maturation in vitro. Trial registration number 0120-546/2018/6

339 EFFECTS OF 17-β ESTRADIOL AND CYSTEAMINE ON IN VITRO MATURATION OF BEEF CATTLE OOCYTES

2010 ◽  
Vol 22 (1) ◽  
pp. 326
Author(s):  
L. Guo ◽  
B. Tang ◽  
X. Ma ◽  
F. Gao ◽  
J. B. Zhang ◽  
...  
Keyword(s):  
Beef Cattle ◽  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Bovine Embryo ◽  
Lh Receptor ◽  
Supplement Group ◽  
Significant Difference ◽  
Maturation Rate ◽  
Harmful Effects

IVM is a critical step in in vitro bovine embryo production. Some materials supplemented in the IVM medium could improve the maturation rate. It was reported that 17-β estradiol (E2) stimulated the FSH-induced follicular growth and expression of the LH receptor in mural granulosa cells, and cysteamine could enhance glutathione synthesis, protect cells against harmful effects caused by oxidative injuries. In order to optimize IVM system of beef cattle oocytes, effects of different supplements (E2 and cysteamine), and dose of supplements on IVM of beef cattle oocytes were investi- gated in this study. The COCs were collected and cultured in the basic IVM media (TCM-199 + 10% FCS + 10 μg mL-1 FSH + 30 μg mL-1 LH) supplemented with hormonesof 0 μg mL-1 (control), 1 μg mL-1, 2 μg mL-1, and 4 μg mL-1 E2, respectively. Statistical analyzes were performed using SPSS (version 9.0) one-way ANOVA. Oocyte maturation rates (mean ± SEM) of four supplement groups were 60.52 ± 1.4%, 74.91 ± 0.25%, 77.25 ± 2.08%, and 62.20 ± 1.87%, respectively. No statistical differences were observed (P > 0.05) between 1 μg mL-1 and 2 μg mL-1 E2 groups; a similar trend was seen between 0 μg mL-1 and 4 μg mL-1 E2 groups. However, oocyte maturation rates (74.91-77.25%) of groups of 1 μg mL-1 and 2 μg mL-1 E2 were significantly higher (P < 0.05) than those (60.52-62.20%) of the other two groups. When basic IVM medium was sup- plemented with 0 μg mL-1 (control), 50 μg mL-1, and 100 μg mL-1 cysteamine, oocyte maturation rates were 75.25 ± 0.25%, 80.56 ± 0.57%, and 76.83 ± 1.82%, respectively. No significant difference (P > 0.05) was observed among them. However, there was approximately five percent higher maturation rate in the supplement group with 50 μg mL-1 cysteamine than those of groups supplemented with 0 μg mL-1 and 100 μg mL-1 cysteamine. Our results indicated: i) the appropriate supplement of E2 (1 μg mL-1 to 2 μg mL-1) could improve in vitro maturation of beef cattle oocytes. Also, the excessive supplement of E2 (>4 μg mL-1) could inhibit the maturation; ii) appropriate supplement of cysteamine (50 μg mL-1) was benefit to the IVM of beef cattle oocytes. This work was supported by the grant from national support plan, China, No. 2007BAD55B03; Corresponding author: Z.Y. Li.

H1foo is essential for in vitro meiotic maturation of bovine oocytes

Zygote ◽  
2014 ◽  
Vol 23 (3) ◽  
pp. 416-425 ◽  
Author(s):  
Yan Yun ◽  
Peng An ◽  
Jing Ning ◽  
Gui-Ming Zhao ◽  
Wen-Lin Yang ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Polar Body ◽  
Linker Histone ◽  
Meiotic Maturation ◽  
Control Group ◽  
Bovine Oocyte ◽  
First Polar Body ◽  
Effective Sirnas ◽  
Bovine Oocytes

SummaryOocyte-specific linker histone, H1foo, is localized on the oocyte chromosomes during the process of meiotic maturation, and is essential for mouse oocyte maturation. Bovine H1foo has been identified, and its expression profile throughout oocyte maturation and early embryo development has been established. However, it has not been confirmed if H1foo is indispensable during bovine oocyte maturation. Effective siRNAs against H1foo were screened in HeLa cells, and then siRNA was microinjected into bovine oocytes to down-regulate H1foo expression. H1foo overexpression was achieved via mRNA injection. Reverse transcription polymerase chain reaction (RT-PCR) results indicated that H1foo was up-regulated by 200% and down-regulated by 70%. Based on the first polar body extrusion (PB1E) rate, H1foo overexpression apparently promoted meiotic progression. The knockdown of H1foo significantly impaired bovine oocyte maturation compared with H1foo overexpression and control groups (H1foo overexpression = 88.7%, H1foo siRNA = 41.2%, control = 71.2%; P < 0.05). This decrease can be rescued by co-injection of a modified H1foo mRNA that has escaped from the siRNA target. However, the H1e (somatic linker histone) overexpression had no effect on PB1E rate when compared with the control group. Therefore we concluded that H1foo is essential for bovine oocyte maturation and its overexpression stimulates the process.

scholarly journals Results of modern tuboperitoneal infertility treatment

2009 ◽  
Vol 66 (1) ◽  
pp. 57-62 ◽  
Author(s):  
Drenka Turjacanin-Pantelic ◽  
Dragana Bojovic-Jovic ◽  
Biljana Arsic ◽  
Eliana Garalejic
Keyword(s):  
Statistical Analysis ◽  
Surgical Treatment ◽  
Infertility Treatment ◽  
P Value ◽  
Modern Approach ◽  
Vitro Fertilization ◽  
Group I ◽  
Significant Difference ◽  
Group Ii

Background/Aim. A modern approach to surgical treatment of tuboperitoneal infertility is based on laporascopic techniques. The aim of this study was to compare results of tuboperitoneal infertility treatment by the use of laparoscopy and classical laparotomy. Methods. A retrospectiveprospective study on 66 women treated operatively form tuboperitoneal infertility was performed. Data from patient's anamnesis and those related to the surgical treatment results, obtained by the use of an inquiry, were used in retrospective and prospective analysis, respectively. Chi-square test was used in statistical analysis. P value < 0.05 was considered significant. Results. Classical laparotomy was used on 34 women in a period from 1996 to 1997, while 32 women were operated laparoscopically in a period from 1999 to 2000. The results were as follows: a total number of conceived women was 16 (24%), seven in the group I (20.6%) and nine in the group II (28.1%); 13 women were with one pregnancy, six in the group I (17.6%) and seven in the group II (22%). Twice pregnant were three women, one in the group I (2.9%) and two in the group II (6.2%). The resulting pregnancies were: five women with abortion spontaneous, two in the group I (5.9%) and three in the group II (9.4%); two women with extrauterine pregnancy in the group I (5.9%); three with pretemporal birth, one in the group I (2.9%) and two in the group II (6.2%), while six women were with the temporal birth, two in the group I (5.9%) and four in the group II (12.5%). Statistical analysis showed that there was no significant difference in the results between these two groups. Conclusion. Surgical treatment of tubeperitoneal infertility, regardless of the used methods (classical laparotomy or laparoscopy) was successful in a great number of women. These methods have a great advantage over in vitro fertilization, and they should not be ignored.

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