Intracellular Ca2+ and antioxidant values induced positive effect on fertilisation ratio and oocyte quality of granulosa cells in patients undergoing in vitro fertilisation

2013 ◽  
Vol 25 (5) ◽  
pp. 746 ◽  
Author(s):  
Esra Nur Tola ◽  
Muhittin Tamer Mungan ◽  
Abdülhadi Cihangir Uğuz ◽  
Mustafa Naziroğlu
Keyword(s):  
Glutathione Peroxidase ◽  
Granulosa Cells ◽  
Oocyte Quality ◽  
Poor Quality ◽  
In Vitro Fertilisation ◽  
Fertilisation Rate ◽  
Gonadotrophin Releasing Hormone ◽  
The Relationship

Oxidative stress is important for promoting oocyte maturation and ovulation within the follicle through calcium ion (Ca2+) influx. The relationship between antioxidant and cytosolic Ca2+ levels and oocyte quality and fertilisation rate in the granulosa cells of patients undergoing in vitro fertilisation was investigated. Granulosa cells were collected from 33 patients. Cytosolic free Ca2+ ([Ca2+]i) concentration, lipid peroxidation, reduced glutathione, glutathione peroxidase and oocyte quality were measured in the granulosa cells. The relationship between two drug protocols was also examined (gonadotrophin-releasing hormone antagonist and agonist protocols) and the same parameters investigated. The [Ca2+]i concentration (P < 0.001), glutathione (P < 0.05) and oocyte quality (P < 0.001) values were significantly higher in the fertilised group than in the non-fertilised group, although glutathione peroxidase activity was significantly (P < 0.05) higher in the non-fertilised group than in the fertilised group. The [Ca2+]i concentrations were also higher (P < 0.001) in the good-quality oocyte groups than in the poor-quality oocyte group. There was no correlation between the two drug protocols and investigated parameters. In conclusion, it was observed that high glutathione and cytosolic Ca2+ concentrations in granulosa cells of patients undergoing in vitro fertilisation tended to increase the fertilisation potential of oocytes.

P–217 The mitochondrial DNA copy number of cumulus granulosa cells is associated with the symmetry of cleavage embryo but not blastocyst quality

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Y Ji ◽  
L Hu
Keyword(s):  
Logistic Regression ◽  
Mitochondrial Dna ◽  
Granulosa Cells ◽  
Copy Number ◽  
Poor Quality ◽  
Dna Copy Number ◽  
Mitochondrial Dna Copy Number ◽  
Fair Poor ◽  
The Relationship

Abstract Study question To study the relationship between mitochondrial DNA copy number of cumulus granulosa cells (CGCs-mtDNA) and the quality of early embryos. Summary answer CGCs-mtDNA was related to the symmetry of cleavage. However, CGCs-mtDNA was not associated with fertilization, blastocysts quality, or blastocysts euploidy. What is known already The potential of early embryonic development mainly depends on the quality of oocytes to a large extent. Mitochondria of CGCs are directly involved in the establishment of oocytes capacitation during oocytes maturation and development. Study design, size, duration This is a retrospective study from December 2018 to January 2019, involving a total of 283 CGCs surrounding Metaphase II oocytes from 49 patients who underwent preimplantation genetic testing for aneuploidy (PGT-A) at the Reproductive and Genetic Hospital of CITIC-Xiangya. Participants/materials, setting, methods We used the TaqMan probes to quantitatively detect mitochondrial DNA copy number of per CGCs by quantitative PCR in mitochondrial genes (MT-ND1 and MT-CO1) and a nuclear gene (β-globin). Besides, according to the nature of the dependent variable, the binary logistic regression model and the logistic regression analysis model of ordered multi-classification were used for multivariate statistical analysis. Main results and the role of chance The CGCs-mtDNA corresponding to fertilized eggs was not different from that of unfertilized eggs in MT-ND1 gene and MT-CO1 gene (MT-ND1: fertilized vs. unfertilized, 600±337 vs. 604±367, P = 0.593; MT-CO1: fertilized vs. unfertilized, 1336±531 vs. 1329±478, P = 0.938). Interestingly, we found that the CGCs-mtDNA of D3 embryos with good quality was statistically higher than that of D3 embryos with fair or poor quality for MT-ND1 gene and MT-CO1 gene (MT-ND1: good quality vs. fair/poor quality, 803±627 vs. 587±307, P = 0.028; MT-CO1: good quality vs. fair/poor quality, 1682±554 vs. 1374±702, P = 0.025). Moreover, the CGCs-mtDNA corresponding to D3 embryos with even cleavage was higher than that of D3 embryos with uneven cleavage (MT-ND1: even cleavage vs. uneven cleavage, 803±627 vs. 590±309, P = 0.036; MT-CO1: even cleavage vs. uneven cleavage, 1562±552 vs. 1316±525, P = 0.037). Besides, we investigated the difference among the CGCs-mtDNA in blastocysts with the good quality, blastocysts with fair or poor quality, and the developmental blocked embryos before the blastocyst stage. But we didn’t found any difference among the above three groups (MT-ND1: P = 0.531; MT-CO1: P = 0.609). In the study of the relationship between CGCs-mtDNA and blastocysts euploidy, we got similar results (MT-ND1: P = 0.602; MT-CO1: P = 0.570). Limitations, reasons for caution The sample size of this study was relatively small. Wider implications of the findings: Although the sample size of this study is limited, our results indicated the importance of mitochondria in CGCs in early embryo development, especially in the first three days. The investigation of mitochondrial function in CGCs may shed light on the mechanism of CGCs-oocyte crosstalk. Trial registration number LL-SC–2019–005

scholarly journals Effect of a high fat diet on ovary morphology, in vitro development, in vitro fertilisation rate and oocyte quality in mice

2015 ◽  
Vol 56 (10) ◽  
pp. 573-579 ◽  
Author(s):  
M Sohrabi ◽  
A Mohammadi Roushandeh ◽  
Z Alizadeh ◽  
A Vahidinia ◽  
M Vahabian ◽  
...  
Keyword(s):  
High Fat Diet ◽  
Oocyte Quality ◽  
In Vitro Fertilisation ◽  
High Fat ◽  
In Vitro Development ◽  
Fertilisation Rate ◽  
Development In Vitro ◽  
Vitro Development

scholarly journals In vitro fertilisation (IVF) versus intracytoplasmic sperm injection (ICSI) in patients without severe male factor infertility: study protocol for the randomised, controlled, multicentre trial INVICSI

BMJ Open ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. e051058
Author(s):  
Sine Berntsen ◽  
Bugge Nøhr ◽  
Marie Louise Grøndahl ◽  
Morten Rønn Petersen ◽  
Lars Franch Andersen ◽  
...  
Keyword(s):  
Live Birth ◽  
Intracytoplasmic Sperm Injection ◽  
Controlled Trial ◽  
Male Partner ◽  
In Vitro Fertilisation ◽  
Male Factor Infertility ◽  
Male Factor ◽  
Fertilisation Rate ◽  
Randomised Controlled

IntroductionOver the last decades, the use of intracytoplasmic sperm injection (ICSI) has increased, even among patients without male factor infertility. The increase has happened even though there is no evidence to support that ICSI results in higher live birth rates compared with conventional in vitro fertilisation (IVF) in cases with nonmale factor infertility. The lack of robust evidence on an advantage of using ICSI over conventional IVF in these patients is problematic since ICSI is more invasive, complex and requires additional resources, time and effort. Therefore, the primary objective of the IVF versus ICSI (INVICSI) study is to determine whether ICSI is superior to standard IVF in patients without severe male factor infertility. The primary outcome measure is first live birth from fresh and frozen-thawed transfers after one stimulated cycle. Secondary outcomes include fertilisation rate, ongoing pregnancy rate, birth weight and congenital anomalies.Methods and analysisThis is a two-armed, multicentre, randomised, controlled trial. In total, 824 couples/women with infertility without severe male factor will be recruited and allocated randomly into two groups (IVF or ICSI) in a 1:1 ratio. Participants will be randomised in variable block sizes and stratified by trial site and age. The main inclusion criteria are (1) no prior IVF/ICSI treatment, (2) male partner sperm with an expected count of minimum 2 million progressive motile spermatozoa following density gradient purification on the day of oocyte pick up and (3) age of the woman between 18 and 42 years.Ethics and disseminationThe study will be performed in accordance with the ethical principles in the Helsinki Declaration. The study is approved by the Scientific Ethical Committee of the Capital Region of Denmark. Study findings will be presented, irrespectively of results at international conferences and submitted for publication in peer-reviewed journals.Trial registration numberNCT04128904. Pre-results.

scholarly journals Mitochondrial enrichment in infertile patients: a review of different mitochondrial replacement therapies

2021 ◽  
Vol 15 ◽  
pp. 263349412110235
Author(s):  
Cristina Rodríguez-Varela ◽  
Sonia Herraiz ◽  
Elena Labarta
Keyword(s):  
Stem Cells ◽  
Genetic Material ◽  
External Source ◽  
Oocyte Quality ◽  
Third Party ◽  
In Vitro Fertilisation ◽  
Mitochondrial Activity ◽  
Mitochondrial Transfer ◽  
Infertile Patients

Poor ovarian responders exhibit a quantitative reduction in their follicular pool, and most cases are also associated with poor oocyte quality due to patient’s age, which leads to impaired in vitro fertilisation outcomes. In particular, poor oocyte quality has been related to mitochondrial dysfunction and/or low mitochondrial count as these organelles are crucial in many essential oocyte processes. Therefore, mitochondrial enrichment has been proposed as a potential therapy option in infertile patients to improve oocyte quality and subsequent in vitro fertilisation outcomes. Nowadays, different options are available for mitochondrial enrichment treatments that are encompassed in two main approaches: heterologous and autologous. In the heterologous approach, mitochondria come from an external source, which is an oocyte donor. These techniques include transferring either a portion of the donor’s oocyte cytoplasm to the recipient oocyte or nuclear material from the patient to the donor’s oocyte. In any case, this approach entails many ethical and safety concerns that mainly arise from the uncertain degree of mitochondrial heteroplasmy deriving from it. Thus the autologous approach is considered a suitable potential tool to improve oocyte quality by overcoming the heteroplasmy issue. Autologous mitochondrial transfer, however, has not yielded as many beneficial outcomes as initially expected. Proposed mitochondrial autologous sources include immature oocytes, granulosa cells, germline stem cells, and adipose-derived stem cells. Presently, it would seem that these autologous techniques do not improve clinical outcomes in human infertile patients. However, further trials still need to be performed to confirm these results. Besides these two main categories, new strategies have arisen for oocyte rejuvenation by improving patient’s own mitochondrial function and avoiding the unknown consequences of third-party genetic material. This is the case of antioxidants, which may enhance mitochondrial activity by counteracting and/or preventing oxidative stress damage. Among others, coenzyme-Q10 and melatonin have shown promising results in low-prognosis infertile patients, although further randomised clinical trials are still necessary.

scholarly journals Sperm binding to ZP2-coated beads improve the efficiency of porcine in vitro fertilisation

Reproduction ◽  
2020 ◽  
Vol 160 (5) ◽  
pp. 725-735
Author(s):  
Julieta Gabriela Hamze ◽  
María Jiménez-Movilla ◽  
Raquel Romar
Keyword(s):  
Acrosome Reaction ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
In Vitro Fertilisation ◽  
Sperm Binding ◽  
Fertilisation Rate ◽  
Gamete Recognition ◽  
Boar Spermatozoa

The role of specific zona pellucida (ZP) glycoproteins in gamete interaction has not yet been elucidated in many species. A recently developed 3D model based on magnetic sepharose beads (B) conjugated to recombinant ZP glycoproteins (BZP) and cumulus cells (CBZP) allows the study of isolated ZP proteins in gamete recognition studies. The objective of this work was to study the role of porcine ZP2, ZP3 and ZP4 proteins in sperm binding, cumulus cell adhesion and acrosome reaction triggering. ZP protein-bound beads were incubated with fresh ejaculated boar spermatozoa and isolated cumulus cells for 24 h. The number of sperm bound to the beads, the acrosomal shrouds (presence of acrosomal content) on the bead’s surface, and the acrosome integrity (by means of PNA-FITC lectin) in bound and unbound sperm were studied. Finally, in vitro matured porcine oocytes mixed with BZP2 were inseminated in vitro using fresh sperm and fertilisation results evaluated. Over 60% of beads had at least one sperm bound after 2 h of coincubation. ZP2-beads (BZP2) and cumulus-ZP2-bead complexes (CBZP2) reached the highest number of sperm per bead, whereas BZP3 and BZP4 models showed the highest number of unbound reacted sperm cells and acrosomal shrouds. Fertilisation efficiency and monospermy rate increased when oocytes were fertilised in the presence of BZP2. We, therefore, conclude that in pigs, it is mainly ZP2 that is involved in sperm-ZP binding whereas ZP3 and ZP4 induce acrosome reaction. Using magnetic sepharose ZP2-bound beads might be a valuable tool to improve the fertilisation rate in pigs.

scholarly journals Embryo transfer as an option to improve fertility in repeat breeder dairy cows

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Arkadiusz Nowicki
Keyword(s):  
Dairy Cows ◽  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Oocyte Quality ◽  
Poor Quality ◽  
Early Embryo ◽  
Reproductive Disorder ◽  
Uterine Environment ◽  
Repeat Breeder

Abstract Repeat breeding is a serious reproductive disorder in dairy cattle. The causes of repeat breeding are multifactorial and there are two main mechanisms: failure of fertilisation or early embryo death, mainly due to poor quality of oocytes and an inadequate uterine environment. Many methods have been used to increase the pregnancy rate for repeat breeder cows, such as intrauterine infusion of antibacterial agents or antibiotics, hormonal treatments for oestrus synchronisation and induction of ovulation, and progesterone supplementation or induction of accessory corpus luteum; however, the results were inconsistent between studies. Embryo transfer (ET) has the capability to minimalise the effects of poor oocyte quality and unfavourable uterine environments on early embryo development during the first seven days after ovulation in repeat breeder cows, and several studies showed that ET significantly improved the pregnancy rate in this group of animals. Thus, ET can be considered an option to increase the conception rate in repeat breeder dairy cows.

scholarly journals Low sperm to egg ratio required for successful in vitro fertilisation in a pair-spawning teleost, Senegalese sole (Solea senegalensis)

2020 ◽  
Author(s):  
Sandra Ramos-Júdez ◽  
Wendy Ángela González-López ◽  
Jhons Huayanay Ostos ◽  
Noemí Cota Mamani ◽  
Carlos Marrero Alemán ◽  
...  
Keyword(s):  
Large Scale ◽  
Reproductive Behaviour ◽  
Poor Quality ◽  
Solea Senegalensis ◽  
In Vitro Fertilisation ◽  
Hatching Rate ◽  
Senegalese Sole ◽  
Single Male ◽  
Close Proximity

AbstractCultured Senegalese sole (Solea senegalensis) breeders fail to spawn fertilised eggs and this bottleneck could be solved with the implementation of large-scale in vitro fertilisation protocols. However, low production of poor-quality sperm has frustrated the development of in vitro fertilisation protocols. Cultured females were induced to ovulate with a 5 µg kg-1 single injection of gonadotropin releasing hormone agonist (GnRHa) and good quality eggs (82.6 ± 9.2% fertilisation) were stripped 41:57 ± 1:46 h after the injection. Sperm was collected from cultured males, diluted in modified Leibovitz and used fresh to fertilise the eggs. A non-linear regression, an exponential rise to a maximum (R = 0.93, P < 0.0001) described the number of motile spermatozoa required to fertilise a viable egg and 1617 motile spermatozoa were sufficient to fertilise 99 ± 12% (± 95% CI) of viable eggs. Similar, spermatozoa egg-1 ratios of 592 ± 611 motile spermatozoa egg-1 were used in large-scale in vitro fertilisations with 190,512 ± 38,471 eggs. The sperm from a single male (145 ± 50 µL or 8.0 ± 6.8 × 108 spermatozoa) was used to fertilise the eggs. The mean hatching rate of the large-scale in vitro fertilisations was 70 ± 14 % to provide 131,540 ± 34,448 larvae per fertilisation. When unfertilised eggs were stored at room temperature the percentage of viable eggs decreased gradually and indicated the sooner eggs were fertilised after stripping the higher the viability of the eggs. The collection of sperm directly into a syringe containing modified Leibovitz significantly increased the percentage of motile spermatozoa (33.4 ± 12.2 %) compared to dilution in modified Leibovitz immediately after collection (6.6 ± 4.9 %). Senegalese sole have a pair-spawning reproductive behaviour characterised by external gamete fertilisation in close proximity with no sperm competition. The low spermatozoa egg-1 ratio required for maximum fertilisation was consistent with this reproductive behaviour and strategy. The provision of a large-scale in vitro fertilisation protocol (200 µL of sperm per 100 mL of eggs) will enable the industry to operate sustainably and implement breeding programs to improve production.

scholarly journals How to Achieve High-Quality Oocytes? The Key Role of Myo-Inositol and Melatonin

2016 ◽  
Vol 2016 ◽  
pp. 1-9 ◽  
Author(s):  
Salvatore Giovanni Vitale ◽  
Paola Rossetti ◽  
Francesco Corrado ◽  
Agnese Maria Chiara Rapisarda ◽  
Sandro La Vignera ◽  
...  
Keyword(s):  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Oocyte Quality ◽  
Fertilization Rate ◽  
In Vitro Models ◽  
The One ◽  
High Level

Assisted reproductive technologies (ART) have experienced growing interest from infertile patients seeking to become pregnant. The quality of oocytes plays a pivotal role in determining ART outcomes. Although many authors have studied how supplementation therapy may affect this important parameter for both in vivo and in vitro models, data are not yet robust enough to support firm conclusions. Regarding this last point, in this review our objective has been to evaluate the state of the art regarding supplementation with melatonin and myo-inositol in order to improve oocyte quality during ART. On the one hand, the antioxidant effect of melatonin is well known as being useful during ovulation and oocyte incubation, two occasions with a high level of oxidative stress. On the other hand, myo-inositol is important in cellular structure and in cellular signaling pathways. Our analysis suggests that the use of these two molecules may significantly improve the quality of oocytes and the quality of embryos: melatonin seems to raise the fertilization rate, and myo-inositol improves the pregnancy rate, although all published studies do not fully agree with these conclusions. However, previous studies have demonstrated that cotreatment improves these results compared with melatonin alone or myo-inositol alone. We recommend that further studies be performed in order to confirm these positive outcomes in routine ART treatment.

scholarly journals Absence of leptin expression and secretion by human luteinized granulosa cells

2003 ◽  
Vol 31 (1) ◽  
pp. 233-239 ◽  
Author(s):  
M Karamouti ◽  
P Kollia ◽  
E Karligiotou ◽  
A Kallitsaris ◽  
N Prapas ◽  
...  
Keyword(s):  
Gene Expression ◽  
Granulosa Cells ◽  
Messenger Rna ◽  
Limit Of Detection ◽  
Culture Media ◽  
Point Of View ◽  
In Vitro Fertilisation ◽  
Ob Gene ◽  
Human Granulosa Cells

Whether leptin is secreted by the human ovary is not known. The available data on leptin gene (ob gene) expression by human granulosa cells are conflicting. The aim of the present study was first to re-examine the expression of leptin messenger RNA (mRNA) by human granulosa cells and second to investigate if these cells have the ability to secrete leptin in cultures. Human luteinized granulosa cells were obtained from normal women undergoing in vitro fertilisation treatment after ovarian stimulation and follicle aspiration. The expression of ob gene was studied by Reverse-Transcriptase Polymerase Chain Reaction (RT-PCR) both in primary granulosa cells treated immediately after oocyte recovery and in cells cultured up to 24 h under baseline and hormonally stimulated conditions (FSH: 100 ng/ml, LH: 100 ng/ml). ob mRNA transcripts were not detected in luteinized granulosa cells, while they were present in adipose tIssue cDNA. Actin gene expression was detected in all studied samples. Using a sensitive radioimmunoassay (lower limit of detection 0.05 ng/ml), leptin was undetectable in the culture media at all points during the 72 h cultures, while at the same time significant amounts of oestradiol and progesterone were produced particularly after the addition of androstendione (1 microM) to the incubation media. These results demonstrate for the first time that leptin is not secreted by human luteinized granulosa cells in cultures. From a physiological point of view, this may contribute to the development of the optimal follicular environment for oocyte maturation during the preovulatory period.

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