14 results on '"Gąsior Ł"'
Search Results
2. Lipid droplet utilization by the mouse embryo
- Author
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Bisogno, S, primary, Arena, R, additional, Fic, K, additional, Gąsior, Ł, additional, and Ptak, GE, additional
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- 2020
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3. Female-age-dependent changes in the lipid fingerprint of the mammalian oocytes.
- Author
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Bisogno S, Depciuch J, Gulzar H, Heber MF, Kobiałka M, Gąsior Ł, Bereta A, Pieczara A, Fic K, Musson R, Garcia Gamero G, Pardo Martinez M, Fornés Pérez A, Tatíčková M, Holubcova Z, Barańska M, and Ptak GE
- Abstract
Study Question: Can oocyte functionality be assessed by observing changes in their intracytoplasmic lipid droplets (LDs) profiles?, Summary Answer: Lipid profile changes can reliably be detected in human oocytes; lipid changes are linked with maternal age and impaired developmental competence in a mouse model., What Is Known Already: In all cellular components, lipid damage is the earliest manifestation of oxidative stress (OS), which leads to a cascade of negative consequences for organelles and DNA. Lipid damage is marked by the accumulation of LDs. We hypothesized that impaired oocyte functionality resulting from aging and associated OS could be assessed by changes in LDs profile, hereafter called lipid fingerprint (LF)., Study Design, Size, Duration: To investigate if it is possible to detect differences in oocyte LF, we subjected human GV-stage oocytes to spectroscopic examinations. For this, a total of 48 oocytes derived from 26 young healthy women (under 33 years of age) with no history of infertility, enrolled in an oocyte donation program, were analyzed. Furthermore, 30 GV human oocytes from 12 women were analyzed by transmission electron microscopy (TEM). To evaluate the effect of oocytes' lipid profile changes on embryo development, a total of 52 C57BL/6 wild-type mice and 125 Gnpat+/- mice were also used., Participants/materials, Setting, Methods: Human oocytes were assessed by label-free cell imaging via coherent anti-Stokes Raman spectroscopy (CARS). Further confirmation of LF changes was conducted using spontaneous Raman followed by Fourier transform infrared (FTIR) spectroscopies and TEM. Additionally, to evaluate whether LF changes are associated with developmental competence, mouse oocytes and blastocysts were evaluated using TEM and the lipid dyes BODIPY and Nile Red. Mouse embryonic exosomes were evaluated using flow cytometry, FTIR and FT-Raman spectroscopies., Main Results and the Role of Chance: Here we demonstrated progressive changes in the LF of oocytes associated with the woman's age consisting of increased LDs size, area, and number. LF variations in oocytes were detectable also within individual donors. This finding makes LF assessment a promising tool to grade oocytes of the same patient, based on their quality. We next demonstrated age-associated changes in oocytes reflected by lipid peroxidation and composition changes; the accumulation of carotenoids; and alterations of structural properties of lipid bilayers. Finally, using a mouse model, we showed that LF changes in oocytes are negatively associated with the secretion of embryonic exosomes prior to implantation. Deficient exosome secretion disrupts communication between the embryo and the uterus and thus may explain recurrent implantation failures in advanced-age patients., Limitations, Reasons for Caution: Due to differences in lipid content between different species' oocytes, the developmental impact of lipid oxidation and consequent LF changes may differ across mammalian oocytes., Wider Implications of the Findings: Our findings open the possibility to develop an innovative tool for oocyte assessment and highlight likely functional connections between oocyte LDs and embryonic exosome secretion. By recognizing the role of oocyte LF in shaping the embryo's ability to implant, our original work points to future directions of research relevant to developmental biology and reproductive medicine., Study Funding/competing Interest(s): This research was funded by National Science Centre of Poland, Grants: 2021/41/B/NZ3/03507 and 2019/35/B/NZ4/03547 (to G.E.P.); 2022/44/C/NZ4/00076 (to M.F.H.) and 2019/35/N/NZ3/03213 (to Ł.G.). M.F.H. is a National Agency for Academic Exchange (NAWA) fellow (GA ULM/2019/1/00097/U/00001). K.F. is a Diamond Grant fellow (Ministry of Education and Science GA 0175/DIA/2019/28). The open-access publication of this article was funded by the Priority Research Area BioS under the program "Excellence Initiative - Research University" at the Jagiellonian University in Krakow. The authors declare no competing interest., Trial Registration Number: N/A., (© The Author(s) 2024. Published by Oxford University Press on behalf of European Society of Human Reproduction and Embryology.)
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- 2024
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4. The antidepressant-like and glioprotective effects of the Y2 receptor antagonist SF-11 in the astroglial degeneration model of depression in rats: Involvement of glutamatergic inhibition.
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Domin H, Konieczny J, Cieślik P, Pochwat B, Wyska E, Szafarz M, Lenda T, Biała D, Gąsior Ł, Śmiałowska M, and Szewczyk B
- Subjects
- Rats, Animals, Astrocytes metabolism, Rats, Sprague-Dawley, Antidepressive Agents pharmacology, Antidepressive Agents metabolism, 2-Aminoadipic Acid pharmacology, Receptors, G-Protein-Coupled metabolism, Prefrontal Cortex metabolism, Depression drug therapy, Depression metabolism, Gliotoxin metabolism, Gliotoxin pharmacology
- Abstract
In this study, we explored the potential antidepressant-like properties of the brain-penetrant Y2 receptor (Y2R) antagonist SF-11 [N-(4-ethoxyphenyl)- 4-(hydroxydiphenylmethyl)- 1-piperidinecarbothioamide] in the astroglial degeneration model of depression with an emphasis on checking the possible mechanisms implicated in this antidepressant-like effect. The model of depression relies on the loss of astrocytes in the medial prefrontal cortex (mPFC) in Sprague-Dawley rats after administering the gliotoxin L-alpha-aminoadipic acid (L-AAA). SF-11 was administered intraperitoneally (i.p.) once (10 mg/kg) or for three consecutive days (10 mg/kg/day), and the effects of L-AAA and SF-11 injected alone or in combination were investigated using the forced swim test (FST), sucrose intake test (SIT), Western blotting, immunohistochemical staining, and microdialysis. SF-11 produced an antidepressant-like effect after single or three-day administration in rats subjected to astrocyte impairment, as demonstrated by the FST and SIT, respectively. Immunoblotting and immunohistochemical analyses showed that SF-11 reversed the L-AAA-induced astrocyte cell death in the mPFC, suggesting it is glioprotective. Microdialysis studies showed that SF-11 decreased extracellular glutamate (Glu) levels compared to basal value when administered alone and compared to the basal value and control group in LAAA-treated rats. The results from immunoblotting analysis indicated the involvement of Y2Rs in the astrocyte ablation model of depression and the antidepressant-like effect of SF-11. In addition, we observed the participation of the caspase-3 apoptotic pathway in the mechanism of gliotoxin action induced by L-AAA. These findings demonstrate that SF-11, a Y2R antagonist, elicited a rapid antidepressant-like response, possibly linked to its ability to inhibit glutamatergic neurotransmission., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)
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- 2024
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5. Long-run real-time PCR analysis of repetitive nuclear elements as a novel tool for DNA damage quantification in single cells: an approach validated on mouse oocytes and fibroblasts.
- Author
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Kotarska K, Gąsior Ł, Rudnicka J, and Polański Z
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- Animals, Mice, Real-Time Polymerase Chain Reaction, Oocytes, Genome, DNA Damage genetics, Fibroblasts
- Abstract
Since DNA damage is of great importance in various biological processes, its rate is frequently assessed both in research studies and in medical diagnostics. The most precise methods of quantifying DNA damage are based on real-time PCR. However, in the conventional version, they require a large amount of genetic material and therefore their usefulness is limited to multicellular samples. Here, we present a novel approach to long-run real-time PCR-based DNA-damage quantification (L1-LORD-Q), which consists in amplification of long interspersed nuclear elements (L1) and allows for analysis of single-cell genomes. The L1-LORD-Q was compared with alternative methods of measuring DNA breaks (Bioanalyzer system, γ-H2AX foci staining), which confirmed its accuracy. Furthermore, it was demonstrated that the L1-LORD-Q is sensitive enough to distinguish between different levels of UV-induced DNA damage. The method was validated on mouse oocytes and fibroblasts, but the general idea is universal and can be applied to various types of cells and species., (© 2023. The Author(s).)
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- 2024
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6. Estrogen fluctuations during the menopausal transition are a risk factor for depressive disorders.
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Turek J and Gąsior Ł
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- Male, Female, Humans, Estrogens, Risk Factors, Serotonin metabolism, Menopause, Depressive Disorder
- Abstract
Women are significantly more likely to develop depression than men. Fluctuations in the ovarian estrogen hormone levels are closely linked with women's well-being. This narrative review discusses the available knowledge on the role of estrogen in modulating brain function and the correlation between changes in estrogen levels and the development of depression. Equally discussed are the possible mechanisms underlying these effects, including the role of estrogen in modulating brain-derived neurotrophic factor activity, serotonin neurotransmission, as well as the induction of inflammatory response and changes in metabolic activity, are discussed., (© 2023. The Author(s).)
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- 2023
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7. Nile Red and BODIPY Staining of Lipid Droplets in Mouse Oocytes and Embryos.
- Author
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Bisogno S, Gąsior Ł, and Ptak GE
- Subjects
- Animals, Boron Compounds, Lipids, Mammals, Mice, Oocytes, Oxazines, Staining and Labeling, Fluorescent Dyes, Lipid Droplets
- Abstract
Lipid droplets (LDs) are intracellular structures composed of hydrophobic lipids. Their amount in oocytes and embryos varies among the mammalian species and even among different strains of the same species. Here we describe a method to stain LDs, which can be applied to previously fixed mouse oocytes and embryos. This method is based on fluorescent dyes, Nile red and BODIPY, which allow visualization and quantification of LDs using conventional and confocal fluorescence microscopy., (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
- Published
- 2023
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8. DNA damage in preimplantation embryos and gametes: specification, clinical relevance and repair strategies.
- Author
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Musson R, Gąsior Ł, Bisogno S, and Ptak GE
- Subjects
- DNA, DNA Damage, Female, Humans, Male, Oocytes physiology, Pregnancy, Blastocyst physiology, Germ Cells
- Abstract
Background: DNA damage is a hazard that affects all cells of the body. DNA-damage repair (DDR) mechanisms are in place to repair damage and restore cellular function, as are other damage-induced processes such as apoptosis, autophagy and senescence. The resilience of germ cells and embryos in response to DNA damage is less well studied compared with other cell types. Given that recent studies have described links between embryonic handling techniques and an increased likelihood of disease in post-natal life, an update is needed to summarize the sources of DNA damage in embryos and their capacity to repair it. In addition, numerous recent publications have detailed novel techniques for detecting and repairing DNA damage in embryos. This information is of interest to medical or scientific personnel who wish to obtain undamaged embryos for use in offspring generation by ART., Objective and Rationale: This review aims to thoroughly discuss sources of DNA damage in male and female gametes and preimplantation embryos. Special consideration is given to current knowledge and limits in DNA damage detection and screening strategies. Finally, obstacles and future perspectives in clinical diagnosis and treatment (repair) of DNA damaged embryos are discussed., Search Methods: Using PubMed and Google Scholar until May 2021, a comprehensive search for peer-reviewed original English-language articles was carried out using keywords relevant to the topic with no limits placed on time. Keywords included 'DNA damage repair', 'gametes', 'sperm', 'oocyte', 'zygote', 'blastocyst' and 'embryo'. References from retrieved articles were also used to obtain additional articles. Literature on the sources and consequences of DNA damage on germ cells and embryos was also searched. Additional papers cited by primary references were included. Results from our own studies were included where relevant., Outcomes: DNA damage in gametes and embryos can differ greatly based on the source and severity. This damage affects the development of the embryo and can lead to long-term health effects on offspring. DDR mechanisms can repair damage to a certain extent, but the factors that play a role in this process are numerous and altogether not well characterized. In this review, we describe the multifactorial origin of DNA damage in male and female gametes and in the embryo, and suggest screening strategies for the selection of healthy gametes and embryos. Furthermore, possible therapeutic solutions to decrease the frequency of DNA damaged gametes and embryos and eventually to repair DNA and increase mitochondrial quality in embryos before their implantation is discussed., Wider Implications: Understanding DNA damage in gametes and embryos is essential for the improvement of techniques that could enhance embryo implantation and pregnancy success. While our knowledge about DNA damage factors and regulatory mechanisms in cells has advanced greatly, the number of feasible practical techniques to avoid or repair damaged embryos remains scarce. Our intention is therefore to focus on strategies to obtain embryos with as little DNA damage as possible, which will impact reproductive biology research with particular significance for reproductive clinicians and embryologists., (© The Author(s) 2022. Published by Oxford University Press on behalf of European Society of Human Reproduction and Embryology.)
- Published
- 2022
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9. Cellular mechanisms ensuring the mammalian female germ cells quality
- Author
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Polański Z and Gąsior Ł
- Subjects
- Animals, Cell Differentiation, Female, Germ Cells, Mammals
- Abstract
Gametes are extremely differentiated cells participating in the fertilization to give the beginning of a new life. Except enabling fertilization, however, the fully functional gamete, should also guarantee full and undisturbed development of the whole individual. The aim of this article is to approximate the mechanisms which occur during mammalian oogenesis which are crucial for ensuring the proper course of development as well as the quality of the genetic material transmitted to the progeny.
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- 2022
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10. Lipid droplets in mammalian eggs are utilized during embryonic diapause.
- Author
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Arena R, Bisogno S, Gąsior Ł, Rudnicka J, Bernhardt L, Haaf T, Zacchini F, Bochenek M, Fic K, Bik E, Barańska M, Bodzoń-Kułakowska A, Suder P, Depciuch J, Gurgul A, Polański Z, and Ptak GE
- Subjects
- Animals, Female, Mice, Blastocyst metabolism, Diapause, Lipid Droplets metabolism, Zygote metabolism
- Abstract
Embryonic diapause (ED) is a temporary arrest of an embryo at the blastocyst stage when it waits for the uterine receptivity signal to implant. ED used by over 100 species may also occur in normally "nondiapausing" mammals when the uterine receptivity signal is blocked or delayed. A large number of lipid droplets (LDs) are stored throughout the preimplantation embryo development, but the amount of lipids varies greatly across different mammalian species. Yet, the role of LDs in the mammalian egg and embryo remains unknown. Here, using a mouse model, we provide evidence that LDs play a crucial role in maintaining ED. By mechanical removal of LDs from zygotes, we demonstrated that delipidated embryos are unable to survive during ED. LDs are not essential for normal prompt implantation, without ED. We further demonstrated that with the progression of ED, the amount of intracellular lipid reduces, and composition changes. This decrease in lipid is caused by a switch from carbohydrate metabolism to lipid catabolism in diapausing blastocysts, which also exhibit increased release of exosomes reflecting elevated embryonic signaling to the mother. We have also shown that presence of LDs in the oocytes of various mammals positively corelates with their species-specific length of diapause. Our results reveal the functional role of LDs in embryonic development. These results can help to develop diagnostic techniques and treatment of recurrent implantation failure and will likely ignite further studies in developmental biology and reproductive medicine fields., Competing Interests: The authors declare no competing interest., (Copyright © 2021 the Author(s). Published by PNAS.)
- Published
- 2021
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11. Mouse single oocyte imaging by MALDI-TOF MS for lipidomics.
- Author
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Bodzon-Kulakowska A, Arena R, Mielczarek P, Hartman K, Kozoł P, Gibuła-Tarlowska E, Wrobel TP, Gąsior Ł, Polański Z, Ptak GE, and Suder P
- Abstract
Reproductive cells are a very special kind of material for the analysis. Depending on the species, their dimensions allow for the application of mass spectrometry imaging-based techniques to receive a reasonable data for interpretation of their condition without any additional sample preparation steps, except for typical sample preparation characteristic for IMS protocols. A comparison between lipid profiles of oocytes could answer the question of the overall quality of the cells in the function of time or conditions of storage. Even tiny differences in the lipid profiles, but still detectable by bioinformatic analysis, could be crucial for the estimation of the conditions of the cells in various stages of development or aging. In our study, MALDI-TOF/TOF MSI was used to analyze and visualize the single oocytes. We deposited the cells on the transparent indium-tin-oxide (ITO) glass and marked their positions, which allowed for the fast localization of the cells and precise laser targeting in the ion source. We also optimized the usage of different MALDI matrices and different approaches. The proposed way of measurement allows analyzing quite a significant quantity of oocytes in a reasonably short time. During the analysis, the lipid composition of the single cell was successfully estimated in a conventional usage of the MALDI ion source, and the localization of lipids was confirmed by imaging mass spectrometry (IMS) analysis. The observed quantity of the lipids allowed for the application of the LIFT™ technique to obtain MS/MS spectra sufficient for lipids' unambiguous identification. We hope that our idea of the oocyte analysis will help to elucidate chemical changes that accompany different processes in which oocytes are involved. There could be such fascinating phenomena as the oocyte maturation, changes in the lipid components during their storage, and much more.
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- 2020
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12. Role of copper in the process of spermatogenesis.
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Ogórek M, Gąsior Ł, Pierzchała O, Daszkiewicz R, and Lenartowicz M
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- Ceruloplasmin, Copper physiology, Humans, Male, Superoxide Dismutase, Trace Elements metabolism, Copper metabolism, Oxidative Stress, Spermatogenesis
- Abstract
Copper (Cu) is an essential trace element required for the normal development of living organisms. Due to its redox potential, copper is a cofactor in many enzymes responsible for important processes in cells. Copper deficiency has a significant influence on the reduction or the total eradication of copper-dependent enzymes in the body, thereby inhibiting cell life processes. On the other hand, copper is a very reactive element and in its free state, it can trigger the production of large amounts of free radicals, which will consequently lead to the damage of proteins and DNA. Because of those reasons, living organisms have developed precise mechanisms regulating the concentration of copper in cells. Copper also plays a very important role in male fertility. It is an essential element for the production of male gametes. The significant role of copper is also described in the processes of cell division - mitotic and meiotic. Copper-dependent enzymes such as ceruloplasmin, superoxide dismutase SOD1 and SOD3, group of metallothionein and cytochrome c oxidase are present at all stages of gametogenesis as well as in the somatic cells of the testis and in the somatic cells of epididymis. Substantial amounts of copper can also be found in liquids associated with sperm in the epididymis and prostate. Copper also affects the integral androgen distribution in terms of fertility on the line hypothalamic-pituitary-testis. Both copper increase and deficiency leads to a significant reduction in male fertility, which spans the entire spectrum of abnormalities at the sperm level, male gonad, production of hormones and distribution of micronutrients such as zinc and iron. Nowadays, the effects of copper on gametes production have become more important and are connected with the increasing levels of pollution with heavy metals in environment.
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- 2017
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13. Mitochondrial functionality in female reproduction.
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Gąsior Ł, Daszkiewicz R, Ogórek M, and Polański Z
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- Aging physiology, Animals, Female, Germ Cells, Humans, Mammals, Oocytes metabolism, Pregnancy, DNA, Mitochondrial, Mitochondria physiology, Reproduction physiology
- Abstract
In most animal species female germ cells are the source of mitochondrial genome for the whole body of individuals. As a source of mitochondrial DNA for future generations the mitochondria in the female germ line undergo dynamic quantitative and qualitative changes. In addition to maintaining the intact template of mitochondrial genome from one generation to another, mitochondrial role in oocytes is much more complex and pleiotropic. The quality of mitochondria determines the ability of meiotic divisions, fertilization ability, and activation after fertilization or sustaining development of a new embryo. The presence of normal number of functional mitochondria is also crucial for proper implantation and pregnancy maintaining. This article addresses issues of mitochondrial role and function in mammalian oocyte and presents new approaches in studies of mitochondrial function in female germ cells.
- Published
- 2017
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14. Prediction of Developmentally Competent Chromatin Conformation in Mouse Antral Oocytes.
- Author
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Daszkiewicz R, Szymoniak M, Gąsior Ł, and Polański Z
- Subjects
- Animals, Cell Nucleolus, Female, Mice, Oogenesis, Ovarian Follicle, Chromatin, Oocytes physiology
- Abstract
Mouse prophase oocytes isolated from antral follicles may possess two alternative types of chromatin configuration: NSN configuration represents more dispersed chromatin and is characteristic mainly for growing oocytes whereas SN configuration, attained upon oocyte growth, comprises more condensed chromatin with a significant fraction concentrated around the nucleolus. Importantly, fully grown oocytes isolated from antral follicles represent a non-homogenous population in which some oocytes posses NSN-type and others SN-type of chromatin conformation. From these two, only oocytes with SN configuration are able to complete full development upon fertilization. We show that among mouse oocytes isolated from antral follicles, those surrounded by cumulus cells were larger and more frequently possessed SN chromatin than oocytes lacking the complete cumulus cell layer. Females primed with PMSG gave a higher number of oocytes with a complete layer of cumulus cells and the frequency of oocytes with SN chromatin was also elevated. Within the whole population of isolated antral oocytes, we observed subtle variation in size which allowed fractionation of oocytes under a stereomicroscope into groups representing oocytes of slightly different sizes. The occurrence of SN chromatin configuration was highly dependent on the oocyte size and its frequency increased gradually in subsequent size groups reaching 95-100% in the group representing the largest oocytes. These findings demonstrate that the subtle differences in the size of antral oocytes allow prediction of the status of their chromatin, thus providing a simple, fast, non-invasive and non-expensive way to select good quality oocytes for ART purposes in mammals.
- Published
- 2016
- Full Text
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