scholarly journals The Presence of Immature GV− Stage Oocytes during IVF/ICSI Is a Marker of Poor Oocyte Quality: A Pilot Study

2020 ◽  
Vol 8 (1) ◽  
pp. 4
Author(s):  
Pia Astbury ◽  
Goutham N. Subramanian ◽  
Jessica Greaney ◽  
Chris Roling ◽  
Jacqui Irving ◽  
...  
Keyword(s):  
Dna Damage ◽  
Pilot Study ◽  
Germinal Vesicle ◽  
Oocyte Retrieval ◽  
Oocyte Quality ◽  
Outcome Data ◽  
Developmental Competence ◽  
Success Rates ◽  
Vitro Fertilization

Here we investigate whether the presence of germinal vesicle-stage oocytes (GV− oocytes) reflects poor oocyte developmental competence (or quality). This was a prospective, non-randomised, cohort pilot-study involving 60 patients undergoing in vitro fertilization/ intracytoplasmic sperm injection for whom complete pregnancy outcome data were available. Patients in whom GV− oocytes were retrieved (GV+) at transvaginal oocyte retrieval (TVOR) were compared with those from whom no GVs were retrieved (GV−). We found that GV+ (n = 29) and GV− (n = 31) patients were similarly aged (35.4 vs. 36.4 years; p = 0.446). GV+ patients had a mean of 2.41 ± 2.03 GVs and comparable yields of MII oocytes to GV− patients (11 ± 6.88 vs. 8.26 ± 4.84; p = 0.077). Compared with GV− patients, GV+ patients had markedly lower implantation rates (11.8% vs. 30.2%; p = 0.022) as well as oocyte utilisation rates for clinical pregnancy (2.3% vs. 6.8%; p = 0.018) and live-birth (1.9% vs. 5.7%; p = 0.029). DNA damage levels measured using γH2AX immunostaining were not different in oocytes from women <36 years versus those ≥36 years (p = 0.606). Thus, patients who have GV− stage oocytes at TVOR exhibit poor oocyte quality reflected in reduced per-oocyte pregnancy success rates and uniformly high levels of oocyte DNA damage.

P–219 mtDNA content in bovine cumulus cells does not predict oocytés developmental competence

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Á Martíne. Moro ◽  
I Lamas-Toranzo ◽  
L González-Brusi ◽  
A Pérez-Gómez ◽  
P Bermejo-Álvarez
Keyword(s):  
Cumulus Cell ◽  
Cumulus Cells ◽  
Oocyte Quality ◽  
Blastocyst Stage ◽  
Early Embryo ◽  
Developmental Competence ◽  
Success Rates ◽  
Developmental Potential ◽  
Mtdna Sequence

Abstract Study question Does cumulus cell mtDNA content correlate with oocyte developmental potential in the bovine model? Summary answer The relative amount of mtDNA content did not vary significantly in oocytes showing different developmental outcomes following IVF What is known already Cumulus cells are closely connected to the oocyte through transzonal projections, serving essential metabolic functions during folliculogenesis. These oocyte-supporting cells are removed and discarded prior to ICSI, thereby constituting an interesting biological material on which to perform molecular analysis aimed to predict oocyte developmental competence. Previous studies have positively associated oocytés mtDNA content with developmental potential in both animal models and women. However, it remains debatable whether mtDNA content in cumulus cells could be used as a proxy to infer oocyte developmental potential. Study design, size, duration Bovine cumulus cells were allocated into three groups according to the developmental potential of the oocyte: 1) oocytes developing to blastocysts following IVF (Bl+Cl+), 2) oocytes cleaving following IVF but arresting their development prior to the blastocyst stage (Bl-Cl+), and 3) oocytes not cleaving following IVF (Bl-Cl-). Relative mtDNA content was analysed in 40 samples/group, each composed by the cumulus cells from one cumulus-oocyte complex (COC). Participants/materials, setting, methods Bovine cumulus-oocyte complexes were obtained from slaughtered cattle and individually matured in vitro (IVM). Following IVM, cumulus cells were removed by hyaluronidase treatment, pelleted, snap frozen in liquid nitrogen and stored at –80 ºC until analysis. Cumulus-free oocytes were fertilized and cultured in vitro individually and development was recorded for each oocyte. Relative mtDNA abundance was determined by qPCR, amplifying a mtDNA sequence (COX1) and a chromosomal sequence (PPIA). Statistical differences were tested by ANOVA. Main results and the role of chance Relative mtDNA abundance did not differ significantly (ANOVA p &gt; 0.05) between the three groups exhibiting different developmental potential (1±0.06 vs. 1.19±0.05 vs. 1.11±0.05, for Bl+Cl+ vs. Bl-Cl+ vs. Bl-Cl-, mean±s.e.m.). Limitations, reasons for caution Experiments were conducted in the bovine model. Although bovine folliculogenesis, monoovulatory ovulation and early embryo development exhibit considerable similarities with that of humans, caution should be taken when extrapolating these data to humans. Wider implications of the findings: The use of molecular markers for oocyte developmental potential in cumulus cells could be used to enhance success rates following single-embryo transfer. Unfortunately, mtDNA in cumulus cells was not found to be a good proxy for oocyte quality. Trial registration number Not applicable

286 DEVELOPMENTAL CAPACITY OF PREPUBERTAL BOVINE OOCYTES CULTURED WITH CYCLIC AMP MODULATORS

2015 ◽  
Vol 27 (1) ◽  
pp. 232 ◽  
Author(s):  
S. M. Bernal ◽  
J. Heinzmann ◽  
D. Herrmann ◽  
U. Baulain ◽  
K.-G. Hadeler ◽  
...  
Keyword(s):  
Oocyte Retrieval ◽  
Developmental Competence ◽  
Vitro Fertilization ◽  
Lactating Cows ◽  
Developmental Capacity ◽  
Commercial Breeding ◽  
Bovine Oocytes ◽  
One Way Anova ◽  
Dmso Vehicle Control

Prepubertal bovine donors are currently used for commercial breeding to accelerate the genetic gain and decrease the generation interval. Nevertheless, it has been reported that their oocyte developmental competence is lower than in adult females. Addition of cAMP regulators during in vitro maturation (IVM) has been suggested to enhance blastocysts rates (Albuz et al. 2010 Hum. Reprod. 25, 2999–3011). Here, we evaluated the effects of the cAMP modulators forskolin, 3-Isobutyl-1-methylxanthine (IBMX), and cilostamide during extended IVM on the developmental capacity of oocytes from prepubertal and adult bovine females. A total of 1851 oocytes from 24 lactating cows (>2 lactations) and 24 prepubertal donors (6–10 mo old) were collected by transvaginal oocyte recovery (OPU) twice per week and divided into 3 experiment groups: (1) TCM24 (OPU medium: PBS; 24 h of IVM; standard protocol/control); (2) cAMP30 [OPU medium: PBS-IBMX (500 μM); 2 h pre-IVM culture using forskolin (100 μM)-IBMX (500 μM) and 30 h of IVM adding cilostamide (20 μM)], and (3) DMSO30 [cAMP modulators are diluted in DMSO)/vehicle control; OPU medium: PBS-DMSO (46.3 mM); 2 h pre-IVM culture (280 mM DMSO) and 30 h IVM (5.6 mM DMSO)]. Following IVM, oocytes were either submitted to in vitro fertilization and embryo culture or fixed in 1% glutaraldehyde at 9, 20, 24, and 30 h after IVM and stained with Hoechst to evaluate their nuclear status. One-way ANOVA was implemented to evaluate recovered oocytes and meiotic stages. The Glimmix procedure from SAS/STAT was performed to compare blastocyst and cleavage rates. Total number of oocytes and IVM-suitable oocytes per donor per OPU session were similar in adult and prepubertal donors (total number/IVM suitable; prepubertal donors: 6.7/4.2, 6.4/4.0, 6.5/3.8; cows: 6.2/4.7, 6.2/4.4, 6.2/4.5 for TCM24, cAMP30 and DMSO30, respectively). At 9 h, cAMP regulators were able to maintain meiotic arrest in prepubertal and adult donors (GV: 80.0 and 40.9%, respectively) compared to standard IVM (GV: 61.1 and 31.2%) and DMSO30 (GV: 40.0 and 26.6%) protocols (P < 0.05). Using the cAMP30 protocol, the percentage of oocytes that reached MII stage at 20 h was lower in adult (4.5%) and prepubertal donors (5.26%) compared to the DMSO30 (50.0 and 42.8%, respectively) and TCM24 (56.2 and 44.4% respectively) protocols. Metaphase II rates after either 24 or 30 h were similar among treatments (prepubertal donors: 88.2, 70.5, and 84.2%; cows: 71.4, 85.7, and 81.2% for TCM24, cAMP30, and DMSO30, respectively; P > 0.05). Cleavage rates (prepubertal donors: 63.4, 54.9, and 52.1%, cows: 56.1, 57.8, and 51.6% for TCM24, cAMP30, and DMSO30, respectively) and blastocysts/presumptive zygotes rates (prepubertal donors: 26.2, 19.6, and 16.2%; cows: 27.5, 28.1, and 21.5% for TCM24, cAMP30, and DMSO30, respectively) did not show significant differences (P > 0.05). Although cAMP modulators delayed the progression through meiosis in adult and prepubertal oocytes, similar blastocysts rates were obtained. Our results suggest so far that oocyte retrieval and competence in prepubertal donors can be similar to that of the adult donors with and without addition of cAMP modulators.

Histone deacetylase inhibitor trichostatin A affects porcine oocyte maturation in vitro

2014 ◽  
Vol 26 (6) ◽  
pp. 806 ◽  
Author(s):  
Yong-Xun Jin ◽  
Ming-Hui Zhao ◽  
Zhong Zheng ◽  
Jung-Suk Kwon ◽  
Seul-Ki Lee ◽  
...  
Keyword(s):  
Trichostatin A ◽  
Germinal Vesicle ◽  
Oocyte Quality ◽  
Histone Deacetylation ◽  
Developmental Competence ◽  
Mitogen Activated Protein Kinase ◽  
Control Group ◽  
Germinal Vesicle Breakdown ◽  
Porcine Oocyte

Previous studies show that porcine oocyte aging resulting from asynchronised IVM impairs embryo developmental competence. In the present study we investigated whether trichostatin A (TSA; an inhibitor of histone deacetylation) prolongs the maturation time and prevents the aging of oocytes. Porcine oocytes were cultured in medium containing increasing concentrations of TSA (300 nM) for 24, 44 or 64 h. The percentage of oocytes that underwent germinal vesicle breakdown was significantly lower in the TSA-treated group (300 nM) than in the control group. TSA did not affect oocyte quality at MII based on levels of maturation-promoting factor, the phosphorylation status of mitogen-activated protein kinase or histone H3K9 acetylation analysis. We also compared the preimplantation developmental competence and the viability of pathenogenetic embryos treated with 100 nM TSA for 24 h and then continuously cultured for another 24 h in TSA free condition. No significant differences were observed for either parameter between the TSA-treated and control groups. These results indicate that TSA prolongs the IVM of porcine oocytes but that oocyte quality and aging are not affected. These findings provide a feasible option by which to adjust the initiation time of downstream experiments based on porcine matured oocytes.

scholarly journals DNA damage in cumulus cells generated after the vitrification of in vitro matured porcine oocytes and its impact on fertilization and embryo development

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Alma López ◽  
Miguel Betancourt ◽  
Yvonne Ducolomb ◽  
Juan José Rodríguez ◽  
Eduardo Casas ◽  
...  
Keyword(s):  
Dna Damage ◽  
Embryo Development ◽  
Tail Length ◽  
Cumulus Cells ◽  
Cell Types ◽  
Oocyte Quality ◽  
Blastocyst Stage ◽  
Developmental Competence ◽  
Development Rates

Abstract Background The evaluation of the DNA damage generated in cumulus cells after mature cumulus-oocyte complexes vitrification can be considered as an indicator of oocyte quality since these cells play important roles in oocyte developmental competence. Therefore, the aim of this study was to determine if matured cumulus-oocyte complexes exposure to cryoprotectants (CPAs) or vitrification affects oocytes and cumulus cells viability, but also if DNA damage is generated in cumulus cells, affecting fertilization and embryo development. Results The DNA damage in cumulus cells was measured using the alkaline comet assay and expressed as Comet Tail Length (CTL) and Olive Tail Moment (OTM). Results demonstrate that oocyte exposure to CPAs or vitrification reduced oocyte (75.5 ± 3.69%, Toxicity; 66.7 ± 4.57%, Vitrification) and cumulus cells viability (32.7 ± 5.85%, Toxicity; 7.7 ± 2.21%, Vitrification) compared to control (95.5 ± 4.04%, oocytes; 89 ± 4.24%, cumulus cells). Also, significantly higher DNA damage expressed as OTM was generated in the cumulus cells after exposure to CPAs and vitrification (39 ± 17.41, 33.6 ± 16.69, respectively) compared to control (7.4 ± 4.22). In addition, fertilization and embryo development rates also decreased after exposure to CPAs (35.3 ± 16.65%, 22.6 ± 3.05%, respectively) and vitrification (32.3 ± 9.29%, 20 ± 1%, respectively). It was also found that fertilization and embryo development rates in granulose-intact oocytes were significantly higher compared to denuded oocytes in the control groups. However, a decline in embryo development to the blastocyst stage was observed after CPAs exposure (1.66 ± 0.57%) or vitrification (2 ± 1%) compared to control (22.3 ± 2.51%). This could be attributed to the reduction in both cell types viability, and the generation of DNA damage in the cumulus cells. Conclusion This study demonstrates that oocyte exposure to CPAs or vitrification reduced viability in oocytes and cumulus cells, and generated DNA damage in the cumulus cells, affecting fertilization and embryo development rates. These findings will allow to understand some of the mechanisms of oocyte damage after vitrification that compromise their developmental capacity, as well as the search for new vitrification strategies to increase fertilization and embryo development rates by preserving the integrity of the cumulus cells.

scholarly journals Oxidative Stress Reduction by Midazolam Premedication during Oocyte Retrieval Procedure: Pilot Study

2021 ◽  
Vol 10 (4) ◽  
pp. 855
Author(s):  
Maja Pešić ◽  
Katarina Kličan-Jaić ◽  
Marinko Vučić ◽  
Krunoslav Kuna ◽  
Andro Košec ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Pilot Study ◽  
In Vitro Fertilization ◽  
Stress Reduction ◽  
Catalase Activity ◽  
Oocyte Retrieval ◽  
Control Group ◽  
Medical Problems ◽  
Vitro Fertilization

Infertility is one of the major medical problems nowadays. Couples who opt for In Vitro Fertilization (IVF) face a great deal of stress which certainly affects the outcome of the procedure. Therefore, we aimed to reduce the stress during the oocyte retrieval procedure by applying midazolam. Total oxidant (TOC) and antioxidant (TAC) capacities of serum, as well as glutathione (GSH) content and catalase activity, were measured in both control and midazolam groups. Follicular fluid was also tested for oxidant capacity and IL1β. Results implied that the midazolam group increased TAC at the end of the procedure. At the same time, the control group decreased GSH at the beginning of the procedure, and both groups decreased catalase activity at the end of the procedure. The results imply that stress during the procedure affects oxidative and antioxidative parameters of the patients, but did not affect the frequency of the pregnancy at the end of this pilot study. Yet, the results imply that oxidative and antioxidative mechanisms during IVF should be investigated in detail as they could affect the outcome of IVF.

scholarly journals Early oocyte triggering followed by in vitro maturation is a good approach in women with resistance ovary syndrome: A case-series

2021 ◽  
Author(s):  
Maryam Eftekhar ◽  
Banafsheh Mohammadi ◽  
Nasim Tabibnejad ◽  
Mohammad Hossein Razi
Keyword(s):  
In Vitro Fertilization ◽  
Intracytoplasmic Sperm Injection ◽  
In Vitro Maturation ◽  
Case Series ◽  
Germinal Vesicle ◽  
Oocyte Retrieval ◽  
Ovary Syndrome ◽  
Vitro Fertilization ◽  
High Doses

Background: Some women represent the inability to respond to endogenous and exogenous gonadotropins during in vitro fertilization/intracytoplasmic sperm injection cycles leading to the follicular developmental arrest. The women with resistant ovaries could benefit from in vitro maturation. Case: This case-series presents pregnancies resulting from initially scheduled conventional in vitro fertilization which led to arrested cycles because of resistant ovary syndrome. The protocol was changed to early oocyte triggering for 15 women due to the small follicles ≤ 12 mm in diameter on day 15 after stimulation with high doses of exogenous gonadotrophins instead of cycle cancellation. Germinal vesicle and metaphase I oocytes that were retrieved from follicles were matured in vitro and inseminated by intracytoplasmic sperm injection. Twenty formed embryos were transferred on day 3 after oocyte retrieval. This resulted in a 30.76% chemical pregnancy out of which no abortion occurred. Therefore, we reported a 30.76% singleton ongoing pregnancy. Conclusion: It seems that early oocyte triggering followed by in vitro maturation may be considered as a good modality in women experiencing follicular resistance to gonadotropins. These cycles can be rescued from cancellation with satisfactory clinical outcomes. Key words: Arrested stimulation cycle, Early oocyte triggering, In vitro maturation, Clinical outcome.

scholarly journals Lipid content, mitochondrial activity and early embryo development in oocyte collected from crossbred cows (bos taurus indicus)

2018 ◽  
Vol 25 (1) ◽  
pp. 120-131
Author(s):  
Yoeli Mendez ◽  
Nohely Parra ◽  
Francisco Baez ◽  
Robert Valeris ◽  
Patricia Villamediana
Keyword(s):  
Lipid Content ◽  
Lipid Droplets ◽  
Oocyte Quality ◽  
Early Embryo ◽  
Developmental Competence ◽  
Mitochondrial Activity ◽  
Vitro Fertilization ◽  
Immature Oocytes ◽  
Crossbred Cows

The objective of this research was to evaluate the effect of phenotypic predominance on lipid content, mitochondrial activity and early developmental competence as indicators of oocyte quality. Cumulus-oocyte complexes (COCs) were recovered through follicular aspiration, and underwent in vitro maturation (IVM), in vitro fertilization (IVF), and in vitro culture (IVC) of presumptive zygotes. Lipid content and mitochondrial activity in immature and IVM oocytes were determined. A maturation rate of 80.6% and 69.3% was found for oocytes predominantly B. indicus and predominantly B. taurus, respectively. Total fertilization rate was 27.6%; 26.1% for predominantly B. indicus oocytes and 29% for predominantly B. taurus oocytes. A total of 55.5% and 57.5% of cleaved embryos after 48 and 72 h post-insemination (hpi) in predominantly B. indicus group were observed, respectively. As for the predominantly B. taurus group, 48.6% and 60.4% of cleaved embryos were found after 48 and 72 hpi, respectively. In both groups, immature oocytes showed a greater amount of small lipidic droplets (p <0.0001); IVM decreased the number of small lipid droplets (p < 0.0001) and increased the number of medium and large lipid droplets (p < 0.0001). Predominantly B. indicus oocytes had a greater number of small and medium-sized lipid droplets, while there were no significant differences in large lipid droplets. IVM oocytes had higher mitochondrial activity than immature oocytes group (p < 0.05) without any effect of phenotypic predominance on this parameter. Assessment of lipid content was not a predictive factor of oocyte quality in crossbred cows.

In vitro Culture of Early Secondary Preantral Follicles to Obtain MII Oocyte Developmental Competence from CD-1 Outbred Mice

2020 ◽  
Author(s):  
Jongwon Kim ◽  
Seungki Lee ◽  
Jung Kyu Choi
Keyword(s):  
In Vitro Culture ◽  
Culture System ◽  
Ovarian Follicle ◽  
Oocyte Quality ◽  
Developmental Competence ◽  
Vitro Fertilization ◽  
Preantral Follicles ◽  
Successful Isolation ◽  
In Vitro Culture System

Background: The ovarian follicle is the fundamental functional tissue unit that consists of mammalian ovary. In humans, it has been known that females are born with a maximum number of follicles or oocytes that are not only non-renewable, but also undergoing degeneration with time with a sharply decreased oocyte quality after the age of 35. Methods: Here, we demonstrate that successful isolation of primary, early secondary and late secondary follicles from the ovaries of CD-1 outbred female mice and in vitro culture system to successfully induce the development of MII oocytes. Result: The 9 days of in vitro culture of early secondary follicles showed significant higher rates in growth and maturation displaying higher numbers of antral follicles and MII oocytes developed from early secondary follicles compared to those cultured for 11 days. However, there was no visible difference induced by the size of initial follicles in the rates of growth and maturation. MII oocytes derived from in vitro culture of early secondary follicles following in vitro fertilization developed into two-cell embryos. These observations demonstrate that developmentally competent MII oocytes can be obtained by in vitro culture of preantral follicles derived from the ovaries of CD-1 mice and reveal a crucial role for CD-1 mice as a novel model for research on human ovarian follicles. Furthermore, this study proposes an in vitro culture system using preantral follicle as a therapeutic strategy for fertility preservation of humans for assisted reproductive medicine. 

Sign in / Sign up

Export Citation Format

Share Document