P–236 The evaluation of apoptosis and luteinization process in cumulus cell culture of IVF patients in terms of embryo development and pregnancy

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
G Dünda. Çiftlik ◽  
M Ergüven ◽  
T İrez
Keyword(s):  
Cell Culture ◽  
Embryo Development ◽  
Cell Apoptosis ◽  
Embryo Quality ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Clinical Pregnancy ◽  
Male Factor ◽  
Apoptosis Rate ◽  
Negative Effect

Abstract Study question Is there any relationship between apoptosis rate and progesterone levels in terms of the degree of embryo quality and clinical pregnancy success? Summary answer The cumulus cell apoptosis and progesterone levels measured at the hCG day were associated with embryo quality and clinical pregnancy success. What is known already The high rate of apoptosis at cumulus cells led to poor embryo development with the clinical pregnancy failure. Study design, size, duration It is prospective study carried out with 40 healthy women diagnosed as the male factor between September 2017 and April 2018. Participants/materials, setting, methods 40 healthy women aged between 27–38 and diagnosed as the male factor who were undergoing IVF participated the study. The cumulus cells were taken after denudation and cultured. The rate of the apoptosis were measured using with TUNEL method every 24 hours for 96 hours. In concominant with the apoptosis evaluation, the embryo progession and clinical pregnancy rate were also monitored. Progesterone levels taken at the hCG day were also measured. Main results and the role of chance In this study, it was shown that cumulus cell apoptosis was lower in the pregnant group compared to the non-pregnant group (p = 0.023), and progesterone value on the hCG day was higher in non-pregnant group (p = 0.021). From the data obtained at the end of the 4th day of culture showed that the apoptosis rate showed a higher tendency in non-pregnant women and this was statistically significant (p = 0.009). In our study, it was found that the ratio of apoptotic cells up to the 4th day showed a positive correlation with the progesterone levels on the hCG day (p = 0.001). Briefly, these results were obtainedfrom this current study that 1) The increase in the rate of apoptotic cells of cumulus cell culture has a negative effect on pregnancy rates, 2) The progesterone levels measured on the day of hCG has a significant negative effect on pregnancy, 3) The increase in progesterone levels acts as a messenger about the apoptosis rate of cumulus cells, 4) The luteinization is strictly associated with the apoptosis of cumulus cells. Limitations, reasons for caution: This was a single center study. Results need to be validated across different centers and high numbered different study population. Wider implications of the findings: The increase in progesterone value acts as a messenger about the rate of apoptosis of cumulus cells may be the prominent indicator to obtain high grade embryo with high clinical pregnancy success. Trial registration number 2017/5–8

10-gingerol induces oxidative stress through HTR1A in cumulus cells: in-vitro and in-silico studies

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Kiptiyah Kiptiyah ◽  
Widodo Widodo ◽  
Gatot Ciptadi ◽  
Aulanni’am Aulanni’Am ◽  
Mohammad A. Widodo ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Culture ◽  
Superoxide Dismutase ◽  
In Silico ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Sod Activity ◽  
In Silico Studies ◽  
Incubation Periods

AbstractBackgroundWe investigated whether 10-gingerol is able to induce oxidative stress in cumulus cells.MethodsFor the in-vitro research, we used a cumulus cell culture in M199, containing 10-gingerol in various concentrations (0, 12, 16, and 20 µM), and detected oxidative stress through superoxide dismutase (SOD) activity and malondialdehyde (MDA) concentrations, with incubation periods of 24, 48, 72, and 96 h. The obtained results were confirmed by in-silico studies.ResultsThe in-vitro data revealed that SOD activity and MDA concentration increased with increasing incubation periods: SOD activity at 0 µM (1.39 ± 0.24i), 12 µM (16.42 ± 0.35ab), 16 µM (17.28 ± 0.55ab), 20 µM (17.81 ± 0.12a), with a contribution of 71.1%. MDA concentration at 0 µM (17.82 ± 1.39 l), 12 µM (72.99 ± 0.31c), 16 µM (79.77 ± 4.19b), 20 µM (85.07 ± 2.57a), with a contribution of 73.1%. Based on this, the in-silico data uncovered that 10˗gingerol induces oxidative stress in cumulus cells by inhibiting HTR1A functions and inactivating GSK3B and AKT˗1.Conclusions10-gingerol induces oxidative stress in cumulus cells through enhancing SOD activity and MDA concentration by inhibiting HTR1A functions and inactivating GSK3B and AKT˗1.

168 INHIBITION OF BOVINE OOCYTE CUMULUS CELL PROGESTERONE SYNTHESIS DURING IN VITRO MATURATION AFFECTS CHROMOSOME ALIGNMENT AND SPINDLE INTEGRITY IN FERTILIZED EGGS AND EARLY EMBRYOS

2014 ◽  
Vol 26 (1) ◽  
pp. 198
Author(s):  
E. Daly ◽  
A. G. Fahey ◽  
M. M. Herlihy ◽  
T. Fair
Keyword(s):  
Embryo Development ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Early Embryo ◽  
Developmental Competence ◽  
Bovine Oocyte ◽  
Spindle Formation ◽  
Vitro Fertilization ◽  
Time Treatment

We have previously demonstrated the importance of progesterone (P4) synthesis by cumulus cells during oocyte maturation in vitro (IVM) for bovine oocyte acquisition of developmental competence and subsequent embryo development (Aparicio et al. 2011 Biol. Reprod. 84). The aim of this study was to identify key processes that may be deregulated by the inhibition of P4 signalling in the cumulus–oocyte complex (COC) during IVM. To this end, good quality immature COC were placed in IVM medium [TCM-199 supplemented with 10% (vol/vol) FCS and 10 ng mL–1 epidermal growth factor] and cultured at 39°C for 22 h in a humidified atmosphere containing 5% CO2, in the presence or absence of 10 μM trilostane (which blocks P4 synthesis by inhibiting 3 β-hydroxysteroid dehydrogenase; Stegram Pharmaceuticals Ltd., Surrey, UK). Matured COC were washed and placed in 250 μL of fertilization medium (25 mM bicarbonate, 22 mM Na-lactate, 1 mM Na-pyruvate, 6 mg mL–1 fatty acid-free BSA, and 10 mg mL–1 heparin). In vitro fertilization (IVF) was performed with 250 μL of frozen–thawed semen at a final concentration of 1 × 106 spermatozoa mL–1 at 39°C under 5% CO2 during 20 h. Presumptive zygotes were denuded, washed, and transferred to 25-μL culture droplets (SOF + 5% FCS) at 39°C under 5% CO2, 90% of N2, and 5% O2 atmosphere with maximum humidity. Subsets of presumptive fertilized eggs and developing embryos were recovered at 6, 72, 120, and 192 h postinsemination (hpi) and processed for confocal whole-mount immunocytochemistry. The meiotic and mitotic spindles and chromosomes were visualised by immunofluorescent labelling of α-tubulin and 4′,6-diamindino-2-phenylindole (DAPI), respectively, and classified as normal if the chromosomes were correctly aligned or appropriately segregated, or abnormal if lagging chromosomes or abnormal chromosome segregation were observed. Samples were collected from 5 replicates (n = 50 zygotes/embryos per treatment, per timepoint) and a total of 157 spindles were observed. Logistic regression analysis was conducted to determine the probability of abnormal spindle formation. The incidence of spindle abnormality was regressed on time, treatment, and treatment by time. For all time points, there was significant reduction in the odds of abnormal spindle formation in control samples versus trilostane-treated samples (P < 0.001). In conclusion, our data imply a role for P4 signalling in maintaining spindle integrity during oocyte meiotic maturation and progression through the initial mitotic divisions of early embryo development in cattle.

Effect of the apoptosis rate observed in oocytes and cumulus cells on embryo development in prepubertal goats

2009 ◽  
Vol 116 (1-2) ◽  
pp. 95-106 ◽  
Author(s):  
B. Anguita ◽  
M.T. Paramio ◽  
R. Morató ◽  
R. Romaguera ◽  
A.R. Jiménez-Macedo ◽  
...  
Keyword(s):  
Embryo Development ◽  
Cumulus Cells ◽  
Apoptosis Rate

scholarly journals Apoptosis in bovine cumulus–oocyte complexes after exposure to polychlorinated biphenyl mixtures during in vitro maturation

Reproduction ◽  
2005 ◽  
Vol 130 (6) ◽  
pp. 857-868 ◽  
Author(s):  
Paola Pocar ◽  
Daniela Nestler ◽  
Michaela Risch ◽  
Bernd Fischer
Keyword(s):  
Cell Apoptosis ◽  
Polychlorinated Biphenyl ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Coplanar Pcbs ◽  
Apoptotic Gene ◽  
Commercial Mixture ◽  
Bovine Oocytes

Aroclor-1254 (A-1254) is a commercial mixture of coplanar (dioxin-like) and non-coplanar (non dioxin-like) polychlorinated biphenyls (PCBs) affecting bovine oocytein vitromaturation (IVM) and developmental competence. In the present study, the role of cumulus cell apoptosis in mediating the toxic effects of PCBs duringin vitromaturation has been investigated. Results indicate that exposure of cumulus–oocyte complexes (COCs) to A-1254 significantly induced apoptosis of cumulus cells. Furthermore, A-1254 significantly increased the expression of the pro-apoptotic gene, Bax, concomitantly reducing the level of the anti-apoptotic gene, Bcl-2, in the cumulus cell compartment. The effects of pure mixtures of coplanar (PCB 77, 126 and 169) or non-coplanar (PCB 52, 101 and 153) PCBs were examined. Exposure of COCs to coplanar PCBs affected maturation at doses as low as 100.6 pg/ml. Furthermore, a significant increase in apoptosis and in Bax mRNA expression was observed. No variations in maturation or apoptosis were observed in the non-coplanar PCB group. To further analyze the role of cumulus cells, COCs and denuded oocytes (DOs) have been exposed to A-1254 or coplanar PCBs during IVM. Exposure of COCs significantly reduced the percentage of matured oocytes after 24 h of culture in both treatments. In contrast, exposure of DOs significantly decreased the maturation rate only at the highest dose investigated (100-fold greater than that affecting COCs). Taken together, the results indicate a direct role of cumulus cell apoptosis in mediating PCB toxicity on bovine oocytes, and a direct relationship between congener planarity and toxicity in bovine oocytes is suggested.

Autologous embryo–cumulus cells co-culture and blastocyst transfer in repeated implantation failures: a collaborative prospective randomized study

Zygote ◽  
2011 ◽  
Vol 20 (2) ◽  
pp. 173-180 ◽  
Author(s):  
M. Benkhalifa ◽  
A. Demirol ◽  
T. Sari ◽  
E. Balashova ◽  
M. Tsouroupaki ◽  
...  
Keyword(s):  
Embryo Development ◽  
Randomized Study ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Prospective Randomized Study ◽  
Human Embryos ◽  
Feeder Cells ◽  
Positive Role ◽  
Significant Difference

SummaryIn repeated implantation failure, the co-culture of human embryos with somatic cells has been reported to promote the improvement of embryos quality, implantation and pregnancy rate. It was reported that feeder cells can be more beneficial to the oocyte and embryo by detoxifying the culture medium and supporting embryo development via different pathways. In this study, 432 patients, each with a minimum of three repeated implantation failures, were accepted for a prospective randomized study with or without autologous cumulus cell embryo co-culture and transfer at day 3 or day 5–6. We also investigated the expression of leukaemia inhibitor factor (LIF) and platelet activating factor receptor (PAF-R) on day 3 confluent cumulus cells. The statistic analysis of the data showed significant difference of implantation and clinical pregnancy rates between classical culture and day 3 compared with co-culture and day 5–6 transfer. The molecular analysis showed that cumulus cells express the LIF and the PAF-R genes and confirmed the possible positive role of growth factors and cytokines in early embryo development. Embryo co-culture systems with autologous cells can be beneficial in routine in vitro fertilization for embryo selection and implantation improvement. More molecular investigations need to be done to improve elucidation of the complex dialogue between the embryo and feeder cells prior to implantation and to understand the involved biological function and molecular process during embryo development.

scholarly journals 252.Oocyte-secreted factor(s) regulate apoptosis of bovine cumulus cells

2004 ◽  
Vol 16 (9) ◽  
pp. 252
Author(s):  
T. S. Hussein ◽  
D. A. Froliand ◽  
J. G. Thompson ◽  
R. B. Gilchrist
Keyword(s):  
Cell Apoptosis ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Dose Effect ◽  
Dependent Manner ◽  
Paracrine Factors ◽  
Cell Proliferation And Differentiation ◽  
Laser Confocal Scanning Microscopy ◽  
Confocal Scanning ◽  
Low Incidence

Paracrine factors secreted by the oocyte affect cumulus cell proliferation and differentiation. These factors may also act in an anti-apoptotic manner, maintaining the low incidence of cellular apoptosis within cumulus cells. The purpose of this study was to determine whether the incidence of apoptosis within cumulus cells is regulated by oocyte-secreted factors (OSF). Bovine cumulus-oocyte complexes (COC) aspirated from abattoir-derived ovaries were randomly allocated to 3 treatments: (1) groups of 5 intact COC; (2) groups of 5 oocytectomised complexes (OOX), where oocytes were removed microsurgically; and (3) groups of 5 OOX co-cultured with 25 denuded oocytes (DO). To examine a dose effect of OSF, OOX were also cultured with increasing numbers of DO (OOX+5DO, OOX+25DO, OOX+50DO). In both experiments, complexes were cultured in their respective treatments for 24 h in 50 �l of oocyte maturation medium (+/– rFSH; 0.1 IU/mL). Apoptosis was assessed using TUNEL, with all nuclei counterstained with propidum iodide (PI). Nikon TE2000 laser confocal scanning microscopy was used to visualise and quantify the incidence of apoptosis (TUNEL/PI). The proportion of apoptotic cells was determined by image analysis. Oocyte removal lead to a significant increase in cumulus cell apoptosis (OOX, 67% apoptotic; COC, 15%; P�<�0.001). However, the incidence of apoptosis in OOX was restored to COC levels when co-cultured with DO (18%; P�>�0.05, COC v. OOX+DO). FSH prevented apoptosis in all treatment groups (P�<�0.05) decreasing the incidence by 27% in OOX and by 12% in COC. Cumulus cell apoptosis within OOX (+/– FSH) was reduced in a dose dependent manner by treating with increasing numbers of DO. These results indicate for the first time that oocyte-secreted factor(s) regulate the incidence of apoptosis within cumulus cells.

scholarly journals The Relationship between Sperm Oxidative Stress Alterations and IVF/ICSI Outcomes: A Systematic Review from Nonhuman Mammals

Biology ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 178
Author(s):  
Jordi Ribas-Maynou ◽  
Marc Yeste ◽  
Albert Salas-Huetos
Keyword(s):  
Oxidative Stress ◽  
Systematic Review ◽  
Embryo Development ◽  
Male Factor ◽  
Fertilization Rates ◽  
Negative Effect ◽  
Infertile Couples ◽  
Icsi Outcomes ◽  
The Relationship

Achieving high embryo quality following IVF and ICSI procedures is a key factor in increasing fertility outcomes in human infertile couples. While the male factor is known to underlie infertility in about 50% of cases, studies performed in human infertile couples have not been able to define the precise effect of sperm affectations upon embryo development. This lack of consistency is, in most cases, due to the heterogeneity of the results caused by the multiple male and female factors that mask the concrete effect of a given sperm parameter. These biases can be reduced with the use of animal gametes, being a good approach for basic researchers to design more homogeneous studies analyzing the specific consequences of a certain affectation. Herein, we conducted a systematic review (March 2020) that assessed the relationship between sperm oxidative stress alterations and IVF/ICSI outcomes in nonhumans mammals. The review was conducted according to PRISMA guidelines and using the MEDLINE-PubMed and EMBASE databases. Thirty articles were included: 11 performed IVF, 17 conducted ICSI, and two carried out both fertilization methods. Most articles were conducted in mouse (43%), cattle (30%) and pig models (10%). After IVF treatments, 80% of studies observed a negative effect of sperm oxidative stress on fertilization rates, and 100% of studies observed a negative effect on blastocyst rates. After ICSI treatments, a positive relationship of sperm oxidative stress with fertilization rates (75% of studies) and with blastocyst rates (83% of studies) was found. In conclusion, the present systematic review shows that sperm oxidative stress is associated with a significant reduction in fertilization rates and in vitro embryo development.

The importance of manganese in the cytoplasmic maturation of cattle oocytes: blastocyst production improvement regardless of cumulus cells presence duringin vitromaturation

Zygote ◽  
2015 ◽  
Vol 24 (1) ◽  
pp. 139-148 ◽  
Author(s):  
Juan Patricio Anchordoquy ◽  
Juan Mateo Anchordoquy ◽  
Matias Angel Sirini ◽  
Juan Alberto Testa ◽  
Pilar Peral-García ◽  
...  
Keyword(s):  
Embryo Development ◽  
Embryo Quality ◽  
Cumulus Cells ◽  
Sod Activity ◽  
Cumulus Expansion ◽  
Production Improvement ◽  
Blastocyst Quality ◽  
Cytoplasmic Maturation

SummaryAdequate dietary intake of manganese (Mn) is required for normal reproductive performance in cattle. This study was carried out to investigate the effect of Mn duringin vitromaturation of bovine cumulus–oocyte complexes (COC) on apoptosis of cumulus cells, cumulus expansion, and superoxide dismutase (SOD) activity in the COC. The role of cumulus cells on Mn transport and subsequent embryo development was also evaluated. Early apoptosis decreased in cumulus cells matured with Mn compared with medium alone. Cumulus expansion did not show differences in COC matured with or without Mn supplementation. SOD activity was higher in COC matured with 6 ng/ml Mn than with 0 ng/ml Mn. Cleavage rates were higher in COC and denuded oocytes co-cultured with cumulus cells, either with or without Mn added toin vitromaturation (IVM) medium. Regardless of the presence of cumulus cells during IVM, the blastocyst rates were higher when 6 ng/ml Mn was supplemented into IVM medium compared with growth in medium alone. Blastocyst quality was enhanced when COC were matured in medium with Mn supplementation. The results of the present study indicated that Mn supplementation to IVM medium enhanced the ‘health’ of COC, and improved subsequent embryo development and embryo quality.

Unravelling the needs of singly in vitro-produced bovine embryos: from cumulus cell co-culture to semi-defined, oil-free culture conditions

2012 ◽  
Vol 24 (8) ◽  
pp. 1084 ◽  
Author(s):  
I. G. F. Goovaerts ◽  
J. L. M. R. Leroy ◽  
A. Langbeen ◽  
E. P. A. Jorssen ◽  
E. Bosmans ◽  
...  
Keyword(s):  
Embryo Development ◽  
Apoptotic Cell ◽  
Specific Interaction ◽  
Cell Monolayer ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Culture Conditions ◽  
In Vitro Production ◽  
Small Surface

Producing bovine in vitro embryos individually is a challenge as it generally leads to impaired embryo development. Earlier research optimised a single embryo in vitro production (IVP) protocol using serum, cumulus cells and oil during culture. As some of these factors are undesirable in certain circumstances, the present study investigated their necessity and possible interactions, and defined their role during single-embryo culture. Although the cumulus cell monolayer produced progesterone, it appeared not to be a key factor in supporting single-embryo development. Because in vitro culture in large medium volumes was shown to impair single-embryo development, two new oil-free culture protocols were tested. Using a 30-µL droplet of medium in 96-well plates with a small surface area resulted in comparable blastocyst rates to those obtained under oil. When serum was used, co-culture with cumulus cells seems necessary, leading to consistently high blastocyst rates. Finally, a serum-free, oil-free culture system using insulin, transferrin, selenium and BSA resulted in embryos with similar total cell numbers and apoptotic cell ratios, but blastocyst rates did not equal those obtained with serum and co-culture. This research additionally stresses the fact that specific interaction mechanisms between somatic cells and a developing in vitro embryo are far from unravelled.

scholarly journals Histone deacetylase inhibitor during in vitro maturation decreases developmental capacity of bovine oocytes

PLoS ONE ◽  
2021 ◽  
Vol 16 (3) ◽  
pp. e0247518
Author(s):  
Thais Preisser Pontelo ◽  
Mauricio Machaim Franco ◽  
Taynan Stonoga Kawamoto ◽  
Felippe Manoel Costa Caixeta ◽  
Ligiane de Oliveira Leme ◽  
...  
Keyword(s):  
Embryo Development ◽  
In Vitro Maturation ◽  
Embryo Quality ◽  
Cumulus Cells ◽  
Developmental Competence ◽  
Nuclear Maturation ◽  
Deacetylase Inhibitor ◽  
Developmental Capacity ◽  
Bovine Oocytes

This study aimed to evaluate the effect of scriptaid during pre-maturation (PIVM) and/or maturation (IVM) on developmental competence of bovine oocytes. Cumulus-oocyte complexes (COCs) were submitted to PIVM for 6 h in the presence or absence of scriptaid. COCs were distributed into five groups: T1-IVM for 22 h, T2-PIVM for 6 h and IVM for 22 h, T3-PIVM with scriptaid for 6 h and IVM for 22 h, T4-PIVM for 6 h and IVM with scriptaid for 22 h, and T5-PIVM with scriptaid for 6 h and IVM with scriptaid for 22 h. Nuclear maturation, gene expression, cumulus cells (CCs) expansion, and embryo development and quality were evaluated. At the end of maturation, all groups presented the majority of oocytes in MII (P>0.05). Only HAT1 gene was differentially expressed (P<0.01) in oocytes with different treatments. Regarding embryo development at D7, T4 (23%) and T5 (18%) had lower blastocyst rate (P<0.05) than the other treatments (T1 = 35%, T2 = 37% and T3 = 32%). No effect was observed when scriptaid in PIVM was used in less competent oocytes (P>0.05). In conclusion, presence of scriptaid in PIVM and/or IVM did not improve developmental competence or embryo quality.

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