Meiotic competence of in vitro grown goat oocytes

Reproduction ◽  
2000 ◽  
pp. 367-373 ◽  
Author(s):  
N Crozet ◽  
M Dahirel ◽  
L Gall
Keyword(s):  
Germinal Vesicle ◽  
Oocyte Diameter ◽  
Germinal Vesicle Breakdown ◽  
Cell Monolayers ◽  
Group A ◽  
The Mean ◽  
Group B ◽  
P34 Cdc2 ◽  
Meiotic Competence

The objective of the present study was to grow meiotically incompetent goat oocytes from early antral follicles in vitro and to render them competent to undergo germinal vesicle breakdown. Cumulus-oocyte complexes with pieces of parietal granulosa cells were isolated from follicles 0.35-0.45 mm in diameter using both mechanical and enzymatic methods. The cumulus-oocyte complexes were divided into two groups according to oocyte diameter (group A: < 95 microm; group B: > 95 microm) and cultured for 8 or 9 days on granulosa cell monolayers. Within 8 days of culture, the mean oocyte diameter increased from 86 +/- 0.4 microm to 95 +/- 0.7 microm in group Aand from 106 +/- 0.2 microm to 109 +/- 0.5 microm in group B. After 9 days of culture, the mean diameter of oocytes from groups A and B were 99 +/- 0.5 microm and 112 +/- 0.4 microm, respectively. The meiotic competence of oocytes grown in vitro was evaluated by in vitro maturation. Within 8 days of culture, only 3% of oocytes from group A and 6% of oocytes from group B acquired the ability to undergo germinal vesicle breakdown. After 9 days of culture, 7% of group A oocytes and 42% of group B oocytes were competent to resume meiosis. The expression of p34(cdc2) in oocytes grown in vitro was analysed by the western blot technique. During 9 days of culture, p34(cdc2) accumulated in both groups of growing oocytes, but its concentration was lower than in fully grown oocytes used as controls. The results showed for the first time that goat oocytes from early antral follicles can grow, accumulate p34(cdc2) and acquire the ability to resume meiosis, when cultured for 9 days on granulosa cell monolayers.

scholarly journals Delay of nuclear maturation and reduction in developmental competence of pig oocytes after mineral oil overlay of in vitro maturation media

Reproduction ◽  
2002 ◽  
pp. 557-564 ◽  
Author(s):  
M Shimada ◽  
N Kawano ◽  
T Terada
Keyword(s):  
Steroid Hormones ◽  
In Vitro Maturation ◽  
Germinal Vesicle ◽  
Mineral Oil ◽  
Developmental Competence ◽  
Cdc2 Kinase ◽  
Germinal Vesicle Breakdown ◽  
High Concentrations ◽  
P34 Cdc2

Steroid hormones, such as progesterone, oestrogen, androgen and meiosis activating sterols, are secreted from cumulus cells that are stimulated by gonadotrophins during maturation of oocytes in vitro. These steroid hormones may be absorbed by mineral oil or paraffin oil; however, in vitro maturation of pig oocytes is commonly performed using medium covered by oil. In this study, high concentrations of progesterone, oestradiol and testosterone were detected in the culture medium after pig cumulus-oocyte complexes (COCs) were cultured with FSH and LH for 44 h in medium without an oil overlay. However, high concentrations of these steroid hormones were not detected in medium when COCs were cultured with the mineral oil overlay. When high concentrations of these steroid hormones were secreted by COCs, germinal vesicle breakdown (GVBD) and the activation of p34(cdc2) kinase and mitogen-activated protein (MAP) kinase in oocytes occurred earlier in comparison with oocytes cultured in medium covered with mineral oil. Moreover, a decrease in p34(cdc2) kinase activity during meiotic progression beyond metaphase I was observed in oocytes cultured in conditions under which high concentrations of steroid hormones were secreted by COCs. In addition, the rate of development to the blastocyst stage after IVF was higher in oocytes matured in medium without an oil overlay. These adverse effects of oil may be explained by absorption by the oil of cumulus-secreted steroids or by the release of toxic compounds into the medium.

scholarly journals Comparative Evaluation of Marginal Fit of Three Provisional Restorative Materials in Different Mediums: An In-Vitro Study

2021 ◽  
Vol 8 (3) ◽  
pp. 9-14
Author(s):  
Manju Choudhary
Keyword(s):  
Methyl Methacrylate ◽  
Room Temperature ◽  
In Vitro Study ◽  
Marginal Adaptation ◽  
Polymerization Shrinkage ◽  
Mean Values ◽  
Group A ◽  
The Mean ◽  
Group B

Temporaries are used as placeholders before the permanent crowns are installed. If the temporary crown is not fitted properly the tooth can be subject to increased decay and gums can become inflamed causing gingivitis which leads to other more serious problems. A provisional fixed restoration will provide a template for defining tooth contour, esthetics, proximal contacts, ridge contacts and occlusion. Margins made by the indirect technique are considered to be more accurate than those made by the direct technique. The purpose of this in vitro study was to compare the marginal accuracy of provisional crowns made from three different biomaterials using the established indirect method in different environments and to evaluate the effect of water absorption on polymerization shrinkage and the effect of polymerization shrinkage occurring in dry storage for a week. Method:Variables used in this study are: a) Revotek LC-light cured composite b) Protemp II c) poly methyl methacrylate. An aluminium master die was machined with dimensions: 5mm length, 10mm gingival diameter, 5 degree taper, and 1mm shoulder then dental stone die was prepared by making an impression of this experimental model using a poly vinyl siloxane material. Both group A and group B consisted of seven specimens each of DPI, ProtempII, Revotek LC. The specimens were evaluated using measuring microscope (Biolux), which had an eyepiece graticule of 1/10th of an mm. The seven specimens in group A (DPI, ProtempII, Revotek LC) were kept in air at room temperature for one week on the dental stone cast and the readings were tabulated and the mean values were obtained. The seven specimens in group B (DPI, ProtempII, and Revotek LC) were kept in water at room temperature for one week and the readings were tabulated in the same manner and the mean values were obtained. Results: Comparative statistics of the mean values of the specimens kept in air at room temperature showed that DPI had the least marginal discrepancies followed by ProtempII, Revotek LC. The values were highly significant (.002) Comparative statistics of the mean values of the specimens kept in water at room temperature showed that DPI had the least marginal discrepancies followed by Revotek LC, ProtempII. The values were highly significant (.009). Conclusion: After one week in air at room temperature and after one week in water at room temperature DPI recorded the minimal marginal discrepancy. When stored in air at room temperature, DPI had the best marginal adaptation. All of the materials showed evidence of continued polymerization shrinkage after storage in air for a week. Water absorption compensated for polymerization shrinkage in DPI and ProtempII whereas Revotek LC was an exception. Keywords: light cured composite, Protemp II-Bis –acryl composite, poly methyl methacrylate, marginal adaptation, provisional restoration.

280 EFFECTS OF HOT SEASON ON BOVINE OOCYTE QUALITY: HOW TO BYPASS THE POOR OOCYTE QUALITY DURING THIS SEASON?

2015 ◽  
Vol 27 (1) ◽  
pp. 229
Author(s):  
L. Boccia ◽  
E. Iacono ◽  
B. Rossi ◽  
B. Merlo
Keyword(s):  
Degradation Products ◽  
Germinal Vesicle ◽  
Oocyte Quality ◽  
Female Reproductive Tract ◽  
Bovine Oocyte ◽  
Germinal Vesicle Breakdown ◽  
Serum Replacement ◽  
Hot Season ◽  
Meiotic Competence

Many authors attribute the decline of reproductive activity in summer to the heat stress, a multifactorial problem in which hyperthermia affects cellular function in various tissues of the female reproductive tract (Hansen et al. 2001; De Rensis et al. 2003). In particular, the combination of high temperatures and high humidity for a long period causes a reduced blood flow to uterus, oviducts, and ovaries, leading to a rise in the concentration of the degradation products of cellular activity. Therefore, the aim of this work was to elucidate the negative effect of the hot season on bovine oocyte quality and evaluate the influence of different factors on the acquisition of meiotic competence. In particular, meiotic competence of bovine oocytes recovered from animals housed at 44°28′00″ N, 11°26′00″ E during spring (March, 4–13°C) and summer (June, 16–27°C) was evaluated. Likewise, in summer the effect of an antioxidant, myo-inositol, the use of serum replacement (SR), and the use of oocytes recovered from cycling heifers (16–18 months) as compared to cows (>24 months) were tested. A total of 1346 abattoir-derived oocytes, equally divided for different experimental groups (over 6 replicates), were in vitro matured in TCM 199 supplemented with EGF (25 ng mL–1), IGF1 (100 ng mL–1), ITS supplement, pFSH-LH (0.1 IU each), and 10% FBS. Myoinositol was added at a concentration of 0, 15, 30, and 50 mM, while 10% SR was used alternatively to FBS. At the end of maturation period (20–22 h), oocytes were denuded and stained with 10 μg mL–1 of Hoechst 33342 at room temperature in the dark. After 15 min they were mounted on glass slides for evaluation of nuclear status using a Nikon Eclipse E400 microscope equipped with fluorescence filters. Nuclear configurations were classified as (a) germinal vesicle (GV), (b) germinal vesicle breakdown (GVBD), (c) metaphase I (M-I), (d) metaphase II (M-II), and (e) degenerated (DEG). Data are expressed as mean ± s.e.m. and were analysed by ANOVA (IBM SPSS Statistics) considering significance at P < 0.05. Oocyte quality of summer oocytes was significantly lower than spring counterparts as result of a higher rate of DEG (8.2 ± 0.6 v. 0.7 ± 0.6) and GV (5.4 ± 0.3 v. 0.4 ± 0.4, respectively; P < 0.05). Myo-inositol supplementation in IVM medium did not significantly affect either oocyte quality or meiotic competence in the hot season, such as the use of SR. When the oocytes were collected from cycling heifers ovaries during summer, the recovery rate of COC/ovary was significantly higher as compared to cows (4.5 v. 2.0), and a lower rate of DEG (1.8 ± 0.2; 8.2 ± 0.6) and GVBD (0.9 ± 0.6; 6.1 ± 0.3) was found (P < 0.05), even if the rate of GV (22.4 ± 0.1 v. 5.4 ± 0.3) was higher (P < 0.05) compared with cow. In conclusion, the hot season negatively affects oocyte quality, myo-inositol does not affect nuclear maturation, and SR can be used alternatively to FBS. The lower age of oocyte donor positively influenced the number of recoverable oocyte and degeneration rate.

scholarly journals A comparison of in vitro fertilization-embryo transfer outcome by two types of soft embryo transfer catheters

2020 ◽  
Vol 7 (3) ◽  
pp. 3678-3685
Author(s):  
Soghra Rabiei ◽  
Mahnaz Yavangi ◽  
Marzieh Farimani ◽  
Iraj Amiri ◽  
Mohamad Fallah ◽  
...  
Keyword(s):  
Clinical Trial ◽  
In Vitro Fertilization ◽  
Embryo Transfer ◽  
Vitro Fertilization ◽  
Significance Level ◽  
Significant Difference ◽  
Group A ◽  
The Mean ◽  
Group B

Introduction: One of the remaining challenges in assisted reproductive procedures, especially in vitro fertilization (IVF), is proper embryo transfer. The aim of this clinical trial was to compare IVFembryo transfer outcome by two types of soft embryo transfer catheters in Hamadan Endometrics and Endometriosis Research Center (Iran). Methods: In this clinical trial study, 100 patients who were candidates for IVF were evaluated. Patients were randomly assigned into two groups (A=50 and B=50). The IVF was identical for both groups until the embryo transfer stage. For group A, soft catheter CH3 PM TRANS SET MINI was used and in group B, KITAZATO soft catheter was used for embryo transfer. All transfers were performed by one person. Patients were recruited using checklists, demographic information, infertility history, beta-human chorionic gonadotropin (ß-hCG) serum levels at day 14 post-transfer, and pregnancy bag 28 days after transfer. The results were analyzed by SPSS software version 16 and using descriptive statistics, chi-square and t-test. The significance level was < 0.05. Results: The mean age of group A and group B was 30.12 and 29.24 years, respectively (p=0.341). The mean duration of infertility in both groups was not statistically significant, and in groups A and B were 4.89 and 4 years, respectively. Ninety % of group A experienced their first IVF experience, while in group B it was slightly lower than 86%, which was not statistically significant. The mean number of eggs obtained in group A was 9.84 and in the group B was 9.88 (p=0.962). The mean number of embryos formed in group A was 6.24 and in group B was 5.72 (p=0.405). There was no statistically significant difference between the two groups in using of Tenaculum, the quality of transmission, and the contamination of the catheter head into the blood or mucus. Conclusion: According to the findings of the present study, the use of KITAZATO catheter compared to PM TRANS SET MINI CH3 catheter for fetal transfer in IVF patients showed no significant difference in pregnancy success rate. However, patients who received the KITAZATO catheter had a slightly higher chance of pregnancy that could be clinically valuable.  

scholarly journals EFFECT OF VAGINAL SILDENAFIL ON IN VITRO FERTILIZATION SUCCESS RATES IN WOMEN WITH PREVIOUS FAILED IN VITRO FERTILIZATION ATTEMPTS

2018 ◽  
Vol 11 (6) ◽  
pp. 486 ◽  
Author(s):  
Ensieh Shahrokh Tehraninejad ◽  
Noushin Khazei ◽  
Elnaz Ayati ◽  
Ali Movafegh ◽  
Omid Azimaraghi
Keyword(s):  
Clinical Trial ◽  
In Vitro Fertilization ◽  
Endometrial Thickness ◽  
Pregnancy Rates ◽  
Control Group ◽  
Vitro Fertilization ◽  
Group A ◽  
The Mean ◽  
Group B

Objectives: Endometrial thickness of <9 mm is a predictor of in vitro fertilization (IVF) failure, although neither pregnancy rates nor the pregnancy outcomes are dependent on the endometrial thickness alone. The impact that uterine artery blood flow has on endometrial growth is dependent on nitric oxide which concentrations could be altered by halting a cyclic guanosine monophosphate-mediated pathway with a phosphodiesterase type 5 selective inhibitor such as sildenafil.Methods: In this clinical trial, 72 patients aged below 45 years which have had at least two earlier failed IVF attempts were randomly split into two groups each consisting of 36 patients. Both groups were started on a long IVF protocol. The case group was also administered 100 mg vaginal sildenafil suppositories daily, starting on day 3 of menstruation which was continued until human chorionic gonadotropin administration. Endometrial thickness was measured using ultrasonography in both groups plus pregnancy rates were assessed in both groups.Results: The mean age of the patients in Group A who received sildenafil; in this clinical trial, 72 patients aged below 45 years which have had at least two previous failed IVF attempts were randomly split into two groups each consisting of 36 patients was 33.8±4.8 in contrast to Group B (control group) with the mean age of 33.8±4.8. Mean endometrial thickness of 8.6±0.1 mm was recorded in Group B compared to 9.0±0.7 mm in Group A (p=0.03). Of all the 36 participants who received sildenafil citrate during the IVF cycle, 12 (33.3%) patients had successful pregnancies while 24 (66.7%) failed to get pregnant. In the control group, out of the 36 participants, 10 (27.8%) patients got pregnant while 26 (72.2%) failed the cycle (p=0.9).Conclusion: This study showed that although using vaginal sildenafil during the IVF cycle does improve endometrial thickness before implantation, this does not necessarily lead to higher pregnancy rates.

Meiotic maturation of bovine oocytes in vitro: improvement of meiotic competence by dibutyryl cyclic adenosine 3′,5′-monophosphate.

1990 ◽  
Vol 68 (4) ◽  
pp. 1182-1187 ◽  
Author(s):  
E. Sato ◽  
M. Matsuo ◽  
H. Miyamoto
Keyword(s):  
Polar Body ◽  
Calf Serum ◽  
Germinal Vesicle ◽  
Razor Blade ◽  
Germinal Vesicle Breakdown ◽  
First Polar Body ◽  
Polar Body Formation ◽  
Bovine Oocytes ◽  
Meiotic Competence

Abstract The present study was undertaken to determine the precise stage of growth at which the ability to resume meiosis is acquired in bovine oocytes. Oocytes of various sizes were isolated from ovaries by mechanical dissection using an 18-gauge needle followed by a razor blade. This method yielded an average of 26.2 ± 7.4 growing and fully grown oocytes from an ovary. Cumulus-enclosed oocytes were cultured in vitro in tissue culture medium 199 containing 10% fetal calf serum. Oocytes ≤ 90 µm in diameter did not resume meiosis. However, germinal vesicle breakdown was observed in oocytes whose diameters exceeded 91 µm. Polar body formation was observed in oocytes with diameters exceeding 101 µm. About 80% of the oocytes with diameters ≥ 121 µm were able to extrude the polar body. The percentage of large oocytes (101 to 120 µm) with first polar body increased when incubated in medium containing dibutyryl cyclic adenosine 3′,5′-monophosphate; however, oocytes 90 to 101 µm did not extrude the first polar body even when cultured in a medium containing dibutyryl cyclic adenosine 3′,5′-monophosphate. These observations indicate that the capability to resume meiosis is acquired gradually during development of oocytes and that dibutyryl cyclic adenosine 3′,5′-monophosphate can improve the meiotic competence of bovine oocytes in culture.

scholarly journals 328MEIOTIC COMPETENCE AND DNA FRAGMENTATION OF PORCINE OOCYTES FROM OVARIES STORED IN VARIOUS TEMPERATURES

2004 ◽  
Vol 16 (2) ◽  
pp. 283 ◽  
Author(s):  
P. Wongsrikeao ◽  
N.W.K. Karja ◽  
A. Budiyanto ◽  
N.R. Mtango ◽  
M. Murakami ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Dna Fragmentation ◽  
Storage Temperature ◽  
Germinal Vesicle ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Germinal Vesicle Breakdown ◽  
C Storage ◽  
Significant Difference ◽  
Meiotic Competence

The aim of this study was to investigate the effects of storage of porcine ovaries at different temperatures before oocyte collection on the nuclear maturation and DNA fragmentation of cumulus-oocyte complexes (COCs). Ovaries were collected at a local abattoir and randomly kept in physiological saline at 4°C, 15°C, 25°C and 35°C. Ovaries were stored for 6 hours prior to follicle aspiration. After storage at each temperature (about 80 oocytes each group), COCs were fixed immediately after aspiration and stained by the terminal deoxynucleotidyl transferase (TdT) nick-end labeling (TUNEL) method to examine the DNA fragmentation under fluorescein microscope. To investigate meiotic competence of the oocytes, some COCs of each treatment group (about 100 oocytes each group) were matured in vitro for 45 hours in a modified North Carolina State University (NCSU)-37 solution supplemented with 10% (v/v) porcine follicular fluid, 0.6mM cysteine, 10IUmL−1 eCG and 10IUmL−1 hCG. After maturation culture, the cumulus cells were removed from COCs and fixed in acetic acid-ethanol (1:3, v/v) for 48–72h. The fixed oocytes were stained with acetic-orcein (1% orcein in 45% acetic acid) and examined under a phase-contrast microscope. Data were subjected to arc-sin transform before analyzing by ANOVA. The proportions of oocytes with DNA fragmentation increased with increasing storage temperature of ovaries (25.2% in 4°C, 31.8% in 15°C, 37.4% in 25°C and 54.7% in 35°C, respectively). There was no significant difference between the proportions of germinal vesicle breakdown (GVBD) of 25°C and 35°C storage groups (74.7 and 83.6%, respectively), but the proportions of 25°C and 35°C storage groups were significantly higher (P&lt;0.05) than those of 4°C and 15°C storage groups (58.1 and 59.6%, respectively). The proportions of oocytes reaching metaphase II (MII) was significantly higher (P&lt;0.05) in the 25°C storage group than in other groups (48.0% in 25°C v. 0% in 4°C, 0% in 15°C and 40.1% in 35°C). Moreover, none of oocytes in 4°C and 15°C storage groups reached MII. These results indicate that 25°C is the most suitable temperature for long-term storage of ovaries to maintain meiotic competence and prevent DNA fragmentation of porcine oocytes.

scholarly journals Comparison of recovery time of ketamine versus nalbuphine used in combination with propofol in females undergoing in-vitro fertilization under intravenous sedation.

2021 ◽  
Vol 28 (02) ◽  
pp. 197-201
Author(s):  
Muazzam Butt ◽  
Saadia Khalique ◽  
Hamza Waheed ◽  
Shumaila Athar ◽  
Zulqarnain Butt ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Recovery Time ◽  
Age Groups ◽  
Intravenous Sedation ◽  
Vitro Fertilization ◽  
Younger Age ◽  
Group A ◽  
The Mean ◽  
Group B

Objective: To compare the mean recovery time after administration of propofol-ketamine versus propofol-nalbuphine combination in patients undergoing in-vitro fertilization procedures under intravenous sedation. Study Design: Randomized control trial. Setting: Department of Anesthesiology, Hameed Latif Hospital, Lahore. Period: 14/11/2016 to 14/11/2017. Material & Methods: 60 Married females of age between 20 to 35 years undergoing elective in-vitro fertilization surgeries of ASA I and II with weight 50-70 kg were included. Group A was treated with nalbuphine and propofol while group B with ketamine and propofol and mean recovery time was calculated. Results: In this study there were total 60 cases, 30 in each group included. The mean age in group A was 27.53±3.36 years while in group B was 26.33±2.44 years. The mean weight in group A was 63.45±3.21 years while in B was 62.78±5.23 years. Recovery time was significantly better in group A (nalbuphine) where it was seen as 4.73±0.98 minutes as compared to 7.17±1.14 minutes in group B with p= 0.01. Recovery time was better in younger age groups (20-27) and again was significantly better in nalbuphine group revealing it in 4.43±0.91 minutes with p= 0.01. Conclusion: The recovery time of nalbuphine is significantly better than ketamine group when combined with propofol.

scholarly journals Kinetic Studies of Soluble Fibrin Complexes in Vivo

1977 ◽  
Author(s):  
F. Schönbach ◽  
I. Mahn ◽  
G. Müller-Berghaus
Keyword(s):  
Kinetic Studies ◽  
Distribution Volume ◽  
Control Group ◽  
Coagulation Disorders ◽  
Soluble Fibrin ◽  
Group A ◽  
The Mean ◽  
Group B

Soluble fibrin complexes may occur in vivo in a variety of coagulation disorders. The aim of this investigation was to elucidate the in vivo behaviour of fibrin-fibrinogen complexes prepared in vitro in a ratio of 1 part fibrin to 20 parts fibrinogen. 12 rabbits (group A) were injected with soluble fibrin complexes containing homologeous I-131-fibrin and I-125-fibrinogen. Another group of 12 rabbits served as control (group B) and received I-131-fibrin solubilized in urea and I-125-fibrinogen separately from each other. Studies were performed over a period of 6 days.The mean distribution volume of fibrin as well as of fibrinogen did not significantly differ between both groups. The elimination characteristics of I-131-fibrin of the soluble fibrin complexes (group A) as well as of the solubilized fibrin (group B) were similiar. The fibrinogen elimination did not differ significantly between the groups: a mean T 1/2 of 47.8 h in group A and a T 1/2 of 46.7 h in group B was calculated.The experiments demonstrate that non-crosslinked soluble fibrin complexes distribute homogeneously in the circulation and dissociate into its subunits. Fibrin is eliminated from the circulating blood without influencing the normal catabolism of fibrinogen.

scholarly journals Chromatin organization and timing of polar body I extrusion identify developmentally competent mouse oocytes

2019 ◽  
Vol 63 (3-4-5) ◽  
pp. 245-251 ◽  
Author(s):  
Federica Cavalera ◽  
Mario Zanoni ◽  
Valeria Merico ◽  
Lucia Sacchi ◽  
Riccardo Bellazzi ◽  
...  
Keyword(s):  
Dna Binding ◽  
Polar Body ◽  
Time Lapse ◽  
Reproductive Technologies ◽  
Developmental Competence ◽  
Cell Stage ◽  
Germinal Vesicle Breakdown ◽  
Group A ◽  
Group B

In the mouse, the use of the DNA-binding fluorochrome Hoechst 33342 allows the classification of fully-grown antral oocytes into two categories distinguished by their chromatin conformation: surrounding nucleolus (SN) and not-surrounding nucleolus (NSN) oocytes, the former capable of completing development, the latter unable to proceed beyond the 2-cell stage. In the present study, time-lapse observation of SN and NSN oocyte GV-to-MII transition highlighted differences in the timing of germinal vesicle breakdown (GVBD) and polar body I (PB-I) extrusion. PB-I extrusion, but not GVBD, revealed the presence of three main groups of significantly different oocytes: Group A (456-576 min) comprising mainly SN oocytes (91.4%), group B (584-728 min) entailing an almost equivalent percentage of SN (52.7%) and NSN (47.3%) oocytes, whereas group C (736-896 min) consisting of almost all NSN (94.4%) oocytes. In a further set of time-lapse experiments, GV oocytes were in vitro matured without Hoechst staining and, depending on the timing of PB-I extrusion, sorted into group A, B or C, inseminated with sperm and observed throughout preimplantation. The results show that 26.2 &plusmn; 12.3% of group A, 2.4 &plusmn; 5.0% of group B and none of group C MII oocytes developed to blastocyst. Overall, this study shows that SN oocytes that complete MI earlier are those with a better developmental competence. The possibility to avoid the use of the invasive DNA-binding fluorochrome Hoechst is relevant for future applications in human and domestic animal reproductive technologies.

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