Dataset Information

Transferrin and antioxidants partly prevented mouse oocyte oxidative damage induced by exposure of cumulus-oocyte complexes to endometrioma fluid.

ABSTRACT:

Background

Exposure of oocytes to the endometrioma fluid has an adverse effect on embryonic quality. To determine whether adding transferrin and antioxidants to culture medium could counteract detrimental effects on mouse cumulus-oocyte complexes (COCs) induced by exposure to endometrioma fluid or not, we conducted an in vitro cross-sectional study using human and mouse COCs.

Methods

Eighteen women who had their oocytes exposed to endometrioma fluid during oocyte retrieval were enrolled. COCs from superovulated ICR female mice were collected. They were first exposed to human endometrioma fluid and then treated by transferrin and/or antioxidants (cysteamine + cystine). Subsequently, COCs function was assessed by molecular methods.

Results

This study observed that human COCs inadvertently exposed to endometrioma fluid in the in vitro fertilization (IVF) group led to a lower good quality embryo rate compared to intracytoplasmic sperm injection (ICSI) group. Exposure of mouse COCs to endometrioma fluid accelerated oocyte oxidative damage, evidenced by significantly reduced CCs viability, defective mitochondrial function, decreased GSH content and increased ROS level, associated with the significantly higher pro-portion of abnormal spindles and lower blastocyst formation (p < 0.05, respectively). This damage could be recovered partly by treating COCs with transferrin and antioxidants (cysteamine + cystine).

Conclusions

Transferrin and antioxidants could reduce the oxidative damage caused by COCs exposure to endometrioma fluid. This finding provides a promising new possibility for intervention in the human oocyte oxidative damage process induced by endometrioma fluid during oocyte pick-up.

SUBMITTER: Ren Z

PROVIDER: S-EPMC7690000 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature

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Publications

Transferrin and antioxidants partly prevented mouse oocyte oxidative damage induced by exposure of cumulus-oocyte complexes to endometrioma fluid.

Ren Zi Z Huang Jiana J Zhou Chuanchuan C Jia Lei L Li Manchao M Liang Xiaoyan X Zeng Haitao H

Journal of ovarian research 20201126 1

<h4>Background</h4>Exposure of oocytes to the endometrioma fluid has an adverse effect on embryonic quality. To determine whether adding transferrin and antioxidants to culture medium could counteract detrimental effects on mouse cumulus-oocyte complexes (COCs) induced by exposure to endometrioma fluid or not, we conducted an in vitro cross-sectional study using human and mouse COCs.<h4>Methods</h4>Eighteen women who had their oocytes exposed to endometrioma fluid during oocyte retrieval were en ...[more]

PMID: 33243289

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Project description:In this study, we tested the overall hypothesis that CC expansion and early embryo development would be improved by including follicular fluid (FF) from small or large follicles in the oocyte maturation medium. In the first experiment, FF aspirated from bovine abattoir ovaries was added to the maturation medium at 0, 25, 50, 75 or 100%. Images of individual COCs were captured at 0, 6, 12 and 19 hours (h) of the maturation period and analyzed to calculate change in the total area over time. Cumulus cell expansion was greatest in COCs matured in 75% and 50% FF, and these differences were detectable at 12 (75% FF only) and 19 h (50% and 75% FF) of maturation. The improvement in CC expansion was greatest when FF from small follicles was used. Treatments for the subsequent experiments were selected based upon the results of the first experiment. Oocyte nuclear maturation rates were observed after supplementing the maturation medium with 0 or 75% FF and maturing for 19 h. The rate of nuclear maturation as determined by the presence or absence of the first polar body was similar between control (0% FF) and treated (75% FF) groups. In the final experiment, COCs were matured in 0%, 50% or 75% FF in preparation for IVF. Duration of the maturation period (12, 19 or 22 h) and size of the follicles from which FF was collected (small or large) also varied. In general, FF supplementation at 50% did not affect the zygotes' developmental potential (neither increased nor decreased). Supplementation of maturation medium with 75% FF from small follicles consistently reduced measures of embryo development while 75% FF from large follicles yielded mixed results. It is concluded that FF supplementation improves CC expansion, but the greater CC expansion does not benefit subsequent embryo development. Notably, however, the 50% FF treatment did not reduce blastocyst rates, indicating that FF can be included in maturation media at concentrations of 50% or less with no detriment to IVF outcomes.

Project description:BackgroundIn vitro embryo production is a highly demanded reproductive technology in horses, which requires the recovery (in vivo or post-mortem) and in vitro maturation (IVM) of oocytes. Oocytes subjected to IVM exhibit poor developmental competence compared to their in vivo counterparts, being this related to a suboptimal composition of commercial maturation media. The objective of this work was to study the effect of different concentrations of secretome obtained from equine preovulatory follicular fluid (FF) on cumulus-oocyte complexes (COCs) during IVM. COCs retrieved in vivo by ovum pick up (OPU) or post-mortem from a slaughterhouse (SLA) were subjected to IVM in the presence or absence of secretome (Control: 0 µg/ml, S20: 20 µg/ml or S40: 40 µg/ml). After IVM, the metabolome of the medium used for oocyte maturation prior (Pre-IVM) and after IVM (Post-IVM), COCs mRNA expression, and oocyte meiotic competence were analysed.ResultsIVM leads to lactic acid production and an acetic acid consumption in COCs obtained from OPU and SLA. However, glucose consumption after IVM was higher in COCs from OPU when S40 was added (Control Pre-IVM vs. S40 Post-IVM: 117.24 ± 7.72 vs. 82.69 ± 4.24; Mean µM ± SEM; p < 0.05), while this was not observed in COCs from SLA. Likewise, secretome enhanced uptake of threonine (Control Pre-IVM vs. S20 Post-IVM vs. S40 Post-IVM: 4.93 ± 0.33 vs. 3.04 ± 0.25 vs. 2.84 ± 0.27; Mean µM ± SEM; p < 0.05) in COCs recovered by OPU. Regarding the relative mRNA expression of candidate genes related to metabolism, Lactate dehydrogenase A (LDHA) expression was significantly downregulated when secretome was added during IVM at 20-40 µg/ml in OPU-derived COCs (Control vs. S20 vs. S40: 1.77 ± 0.14 vs. 1 ± 0.25 vs. 1.23 ± 0.14; fold change ± SEM; p < 0.05), but not in SLA COCs.ConclusionsThe addition of secretome during in vitro maturation (IVM) affects the gene expression of LDHA, glucose metabolism, and amino acid turnover in equine cumulus-oocyte complexes (COCs), with diverging outcomes observed between COCs retrieved using ovum pick up (OPU) and slaughterhouse-derived COCs (SLA).

Project description:Study questionIs the abundance of certain biochemical compounds in human cumulus cells (CCs) related to oocyte quality?Summary answerMalonate, 5-oxyproline, and erythronate were positively associated with pregnancy potential.What is known alreadyCCs 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. Mitochondrial DNA content and transcriptional analyses in CC have been shown to provide a poor predictive value of oocyte competence, but the untargeted analysis of biochemical compounds (metabolomics) has been unexplored.Study design, size, durationCCs were obtained from three groups of cumulus-oocyte complexes (COCs) of known developmental potential: oocytes not developing to blastocyst following ICSI (Bl-); oocytes developing to blastocyst but failing to establish pregnancy following embryo transfer (P-); and oocytes developing to blastocyst able to establish a pregnancy (P+). Metabolomics analyses were performed on 12 samples per group, each sample comprising the CC recovered from a single COC.Participants/materials, setting, methodsHuman CC samples were obtained from IVF treatments. Only unfrozen oocytes and embryos not submitted to preimplantation genetic testing were included in the analysis. Metabolomics analysis was performed by ultra-high performance liquid chromatography-tandem mass spectroscopy.Main results and the role of chanceThe analysis identified 98 compounds, five of which were differentially abundant (P < 0.05) between groups: asparagine, proline, and malonate were less abundant in P- compared to Bl-, malonate and 5-oxoproline were less abundant in P- group compared to P+, and erythronate was less abundant in Bl- group compared to P+. No significant association between the abundance of the compounds identified and donor age or BMI was noted.Limitations, reasons for cautionData dispersion and the lack of coherence between developmental groups preclude the direct use of metabolic markers in clinical practice, where the uterine environment plays a major role in pregnancy outcome. The abundance of other compounds not detected by the analysis may be associated with oocyte competence. As donors were lean (only two with BMI > 30 kg/m2) and young (<34 years old), a possible effect of obesity or advanced age on the CC metabolome could not be determined.Wider implications of the findingsThe abundance of malonate, 5-oxyproline, and erythronate in CC was significantly higher in COCs ultimately establishing pregnancy, providing clues on the pathways required for oocyte competence. The untargeted analysis uncovered the presence of compounds that were not expected in CC, such as β-citrylglutamate and the neurotransmitter N-acetyl-aspartyl-glutamate, which may play roles in chromatin remodeling and signaling, respectively.Study funding/competing interest(s)Research was supported by the Industrial Doctorate Project IND2017/BIO-7748 funded by Madrid Region Government. The authors declare no competing interest.Trial registration numberN/A.

Dataset Information

Transferrin and antioxidants partly prevented mouse oocyte oxidative damage induced by exposure of cumulus-oocyte complexes to endometrioma fluid.

Background

Methods

Results

Conclusions

Publications

Transferrin and antioxidants partly prevented mouse oocyte oxidative damage induced by exposure of cumulus-oocyte complexes to endometrioma fluid.

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