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Keywords = in vivo matured oocytes

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17 pages, 1750 KiB  
Review
Reproductive Challenges in Ruminants Under Heat Stress: A Review of Follicular, Oocyte, and Embryonic Responses
by Danisvânia Ripardo Nascimento, Venância Antonia Nunes Azevedo, Regislane Pinto Ribeiro, Gabrielle de Oliveira Ximenes, Andreza de Aguiar Silva, Efigênia Cordeiro Barbalho, Laryssa Gondim Barrozo, Sueline Cavalcante Chaves, Maria Samires Martins Castro, Erica Costa Marcelino, Leopoldo Rugieri Carvalho Vaz da Silva, André Mariano Batista and José Roberto Viana Silva
Animals 2025, 15(15), 2296; https://doi.org/10.3390/ani15152296 - 6 Aug 2025
Abstract
This review aims to discuss how heat stress affects ovarian follicles and oocytes, steroidogenesis, and embryo development in ruminants. The literature shows that quiescent primordial follicles appear to be less susceptible to heat stress, but from the primary follicle stage onwards, they begin [...] Read more.
This review aims to discuss how heat stress affects ovarian follicles and oocytes, steroidogenesis, and embryo development in ruminants. The literature shows that quiescent primordial follicles appear to be less susceptible to heat stress, but from the primary follicle stage onwards, they begin to suffer the consequences of heat stress. These adverse effects are exacerbated when the follicles are cultured in vitro. In antral follicles, heat stress reduces granulosa cell viability and proliferation in both in vivo and in vitro models. Oocyte maturation, both nuclear and cytoplasmic, is also compromised, and embryo quality declines under elevated thermal conditions. These effects are linked to intracellular disturbances, including oxidative imbalance, mitochondrial dysfunction, and altered hormonal signaling. The differences between in vivo and in vitro responses reflect the complexity of the biological impact of heat stress and emphasize the protective role of the physiological microenvironment. A better understanding of how heat stress alters the function of ovarian follicles, oocytes, and embryos is crucial. This knowledge is critical to devise effective strategies that mitigate damage, support fertility, and improve outcomes in assisted reproduction for livestock exposed to high environmental temperatures. Full article
(This article belongs to the Special Issue Heat Stress in Animal Oocytes: Impacts, Evaluation, and Alleviation)
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22 pages, 1986 KiB  
Review
Curcumin as Therapeutic Modulator of Impaired Antioxidant Defense System: Implications for Oxidative Stress-Associated Reproductive Dysfunction
by Tuba Latif Virk, Qi Liu, Yuguo Yuan, Xianyu Xu and Fenglei Chen
Biology 2025, 14(7), 750; https://doi.org/10.3390/biology14070750 - 23 Jun 2025
Viewed by 539
Abstract
One of the critical challenges in assisted reproductive technology (ART) is the inadequacy of effective regulation of reactive oxygen species. Simultaneously, the endogenous antioxidant defense system plays a significant role in combating oxidative stress across various physiological stages of embryonic development. However, these [...] Read more.
One of the critical challenges in assisted reproductive technology (ART) is the inadequacy of effective regulation of reactive oxygen species. Simultaneously, the endogenous antioxidant defense system plays a significant role in combating oxidative stress across various physiological stages of embryonic development. However, these intrinsic defense systems alone are insufficient as they rely on exogenous antioxidants that interact synergistically to enhance and sustain antioxidant capacity. Considering the principal role of antioxidants in mitigating oxidative stress in oocyte growth, identifying reliable and non-toxic antioxidants is an essential prerequisite for effective therapeutic applications. Thus, owing to the need to explore exogenous antioxidants, we attempted to summarize and analyze the literature data defining the potential use of curcumin in mitigating oxidative stress to promote oocyte maturation through in vivo and in vitro model studies. Recent studies demonstrated the protective role of curcumin against oxidative stress and the inflammatory response, primarily through the upregulation of key antioxidant enzymes (including SOD, CAT and GPx), a reduction in oxidative stress markers (e.g., ROS, MDA) and by suppressing the pro-inflammatory signaling pathways (such as NF-kB, JAK/STAT) while activating the NRF2/HO-1 pathway to further enhance the cellular antioxidant defense. Advancing curcumin as a therapeutic agent necessitates a thorough understanding of curcumin’s molecular mechanisms and targeted pharmacological effectiveness to treat female infertility, and despite the progress in enhancing curcumin’s bioavailability, the optimal dosing strategies still need to be defined. Future studies are required to develop strategies to augment antioxidant defense mechanisms (modeling in vivo and in vitro studies) using curcumin with a specific emphasis on curcumin’s role in improving mitochondrial activity. This approach is expected to represent a significant advancement in the field of medicine, offering novel therapeutic possibilities. Full article
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19 pages, 2694 KiB  
Article
Biphasic CAPA-IVM Improves Equine Oocyte Quality and Subsequent Embryo Development Without Inducing Genetic Aberrations
by Muhammad Fakhar-I-Adil, Daniel Angel-Velez, Emin Araftpoor, Qurratul Ain Amin, Mohamed Hedia, Marcel Bühler, Kris Gevaert, Björn Menten, Ann Van Soom, Susana Marina Chuva de Sousa Lopes, Dominic Stoop, Chloë De Roo, Katrien Smits and Björn Heindryckx
Int. J. Mol. Sci. 2025, 26(12), 5495; https://doi.org/10.3390/ijms26125495 - 8 Jun 2025
Viewed by 937
Abstract
In vitro maturation (IVM) of oocytes retrieved from ovum pick-up (OPU) or ovarian tissue (OT) is a standard approach for patients with specific conditions where prior hormonal stimulation is contraindicated. However, the developmental competence of oocytes matured in vitro is still inferior to [...] Read more.
In vitro maturation (IVM) of oocytes retrieved from ovum pick-up (OPU) or ovarian tissue (OT) is a standard approach for patients with specific conditions where prior hormonal stimulation is contraindicated. However, the developmental competence of oocytes matured in vitro is still inferior to that of oocytes matured in vivo. Capacitation IVM (CAPA-IVM) includes an extra step of pre-maturation culture (PMC) with c-type natriuretic peptide (CNP) as a meiotic arrestor to better synchronize cytoplasmic and nuclear maturity in oocytes by allowing the cytoplasm additional time to acquire essential components critical for optimal competency. This study aims to evaluate the effect of CAPA-IVM on equine oocyte quality and developmental competence. Immature cumulus–oocyte complexes (COCs) were retrieved from slaughterhouse ovaries and matured in vitro either in CAPA-IVM (short 6 h, long 24 h pre-maturation) or standard IVM. Mature oocytes from each group were analyzed for calcium-releasing potential (n = 52) and single-oocyte proteomics (n = 44), and embryo development (n = 229) was assessed after fertilization with piezo-drilled intracytoplasmic sperm injection (ICSI). Genetic analysis of developed blastocysts (n = 41) was performed to detect chromosomal aberrations. Our findings demonstrate that CAPA-IVM of equine COCs yields significantly higher maturation rates than controls. Moreover, short CAPA-IVM with six hours pre-maturation culture showed substantially higher embryo development potential than the control group (20/69 vs. 9/63, respectively). Genetic analysis revealed a high euploidy rate in equine blastocysts regardless of the maturation conditions. Live calcium imaging of the fertilized oocytes demonstrated that the majority of oocytes displayed non-continuous calcium oscillation patterns, irrespective of maturation conditions. Single-oocyte proteomics reveals a comparable proteomic landscape between mature oocytes subjected to short CAPA-IVM and standard IVM. However, we identified four enriched gene sets with positive enrichment scores after short CAPA-IVM, related to cytoskeleton regulation, ribosomal function, and cytosolic components. Our findings indicate that CAPA-IVM holds the potential to improve oocyte quality and competence in horses. However, further fine-tuning of culture conditions would benefit the effective use of these IVM systems. Moreover, given that the mare serves as an excellent model for human reproduction, the molecular trends identified in this study could provide valuable insights for advancing human artificial reproductive technologies. Full article
(This article belongs to the Special Issue Molecular Research on Embryo Developmental Potential)
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28 pages, 3298 KiB  
Article
Expression Profiling and Function Analysis Identified New Cumulus Cells-Expressed Genes and miRNAs Predictive of Oocyte Developmental Potential
by Min Zhang, Meng-Meng Wang, Fa-Li Zhang, Nan He, Ming-Jiu Luo, Shuai Gong, Fu-Yin Fu, Hong-Jie Yuan, Jie Zhang and Jing-He Tan
Cells 2025, 14(11), 791; https://doi.org/10.3390/cells14110791 - 28 May 2025
Viewed by 548
Abstract
Although prior studies have identified cumulus cells (CCs)-expressed genes and miRNAs that regulate cumulus expansion and/or CC apoptosis and may serve as markers for selecting competent oocytes and embryos, there remains an urgent need to identify CCs-expressed genes and miRNAs whose expression levels [...] Read more.
Although prior studies have identified cumulus cells (CCs)-expressed genes and miRNAs that regulate cumulus expansion and/or CC apoptosis and may serve as markers for selecting competent oocytes and embryos, there remains an urgent need to identify CCs-expressed genes and miRNAs whose expression levels are directly correlated with oocyte developmental potential (DP). In this study, we first established CC models from mouse cumulus-oocyte complexes (COCs) that exhibited significantly different DP following in vitro or in vivo maturation. Subsequently, we performed mRNA/miRNA sequencing and functional analyses using these in vitro and in vivo CC models. We identified and validated Spp1, Fn1, Sdc1, and Ngf as DP-beneficial genes; Fos and Jun as DP-detrimental genes; and miR-7686-5p, miR-133a-3p, novel-miR-239, novel-miR-193, and miR-339-5p as DP-detrimental miRNAs. Finally, by employing a well-in-well activation/embryo culture system that enables tracking the COC origin of CCs and embryos, we further validated Spp1 and Ngf as DP-beneficial genes, Jun as the DP-detrimental gene, and miR-7686-5p, novel-miR-239, and miR-339-5p as DP-detrimental miRNAs. In conclusion, we identified and validated new sets of CCs-expressed genes and miRNAs whose expression levels were directly correlated with oocyte DP. These newly identified genes and miRNAs may serve as potential biomarkers for selecting competent oocytes and embryos. Full article
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13 pages, 1806 KiB  
Article
Ovarian Stromal Cell-Conditioned Media, but Not Co-Culture, Improves Survival in Feline Follicles
by Batsheva Marks, Jennifer Beth Nagashima, Carol L. Keefer and Nucharin Songsasen
Animals 2025, 15(11), 1539; https://doi.org/10.3390/ani15111539 - 24 May 2025
Viewed by 413
Abstract
Ovarian stromal cells play roles in in vivo folliculogenesis; however, little is known about their effect on in vitro cultured follicles. This study investigated the impact of ovarian stromal cell co-culture or conditioned medium (CM) on the survival and development of domestic cat [...] Read more.
Ovarian stromal cells play roles in in vivo folliculogenesis; however, little is known about their effect on in vitro cultured follicles. This study investigated the impact of ovarian stromal cell co-culture or conditioned medium (CM) on the survival and development of domestic cat follicles in vitro. Preantral (n = 148 follicles), early antral (n = 92), and antral (n = 22) stage cat follicles were divided into five groups (control, ovarian stromal cell co-culture, 20% CM, 50% CM, and 100% CM), cultured for 13 days, and evaluated for survival, growth, and the mRNA expression of CYP19A, GDF9, and FSHR. Additional follicles (n = 199) were isolated, divided into three groups (control, co-culture, and 100% CM), cultured for 10 days, and oocytes were subjected to in vitro maturation (IVM). More follicles (p ≤ 0.01) cultured in 100% CM survived until day 11 of culture than other groups. Antral follicle survival was significantly lower than pre- or early antral (p ≤ 0.0001). However, no differences (p > 0.05) in growth were detected across the treatments. CYP19A expression was upregulated (p ≤ 0.001) in the 50% CM-treated follicles. Furthermore, no differences (p > 0.05) were found in IVM rates between cultures. In summary, the findings demonstrate that conditioned medium collected from primary cultures of ovarian stromal cells improves in vitro survival of isolated cat follicles. Full article
(This article belongs to the Section Animal Reproduction)
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18 pages, 833 KiB  
Article
In Vitro Maturation of Bovine Oocytes in the Presence of Resveratrol and Ellagic Acid but Not Chlorogenic Acid Modulates Blastocyst Antioxidant Gene Expression Without Impacting Embryo Development and Oxygen Consumption
by Katrin Giller, Dominique Schmid, Idil Serbetci, Manuel Meleán, Sarah Greve, Ferdinand von Meyenn, Heinrich Bollwein and Carolina Herrera
Antioxidants 2025, 14(6), 621; https://doi.org/10.3390/antiox14060621 - 23 May 2025
Viewed by 654
Abstract
In vitro fertilization is used to produce embryos from high-genetic-merit cattle. However, these embryos often exhibit inferior quality than those derived in vivo, possibly due to increased oxidative stress. This study investigates whether adding antioxidant polyphenols (resveratrol (RV), chlorogenic acid (CA), ellagic acid [...] Read more.
In vitro fertilization is used to produce embryos from high-genetic-merit cattle. However, these embryos often exhibit inferior quality than those derived in vivo, possibly due to increased oxidative stress. This study investigates whether adding antioxidant polyphenols (resveratrol (RV), chlorogenic acid (CA), ellagic acid (EA)) to the in vitro maturation (IVM) medium at 0.25, 0.5, and 1 μM could improve embryo development. Oxygen consumption and gene expression were evaluated at the blastocyst stage following treatment with 1 μM of each polyphenol. Embryo development (cleavage, blastocyst, and hatched blastocyst rates) and oxygen consumption were not significantly affected by polyphenols. However, RV significantly upregulated the mRNA expression of the antioxidant enzyme glutathione peroxidase 4 (GPX4), while GPX4 expression was significantly downregulated by EA. Expression of other gene markers related to antioxidant defense, apoptosis, development, and metabolism was not significantly affected. The results indicate that applying RV, CA, and EA during bovine oocyte IVM does not enhance in vitro embryo development at the tested concentrations. Given the opposing effects of RV and EA on the expression of GPX4, the effects of those polyphenols regarding the protection of embryos from oxidative stress and potential long-term effects on the offspring remain to be elucidated. Full article
(This article belongs to the Special Issue Redox Regulation in Animal Reproduction)
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20 pages, 13817 KiB  
Article
Methylmercury Chloride Exposure Affects Oocyte Maturation Through AMPK/mTOR-Mediated Mitochondrial Autophagy
by Shengkui Hou, Caiyu Wang, Xin Ma, Jing Zhao, Jun Wang, Yi Fang, Hongyu Liu, He Ding, Jing Guo and Wenfa Lu
Int. J. Mol. Sci. 2025, 26(8), 3603; https://doi.org/10.3390/ijms26083603 - 11 Apr 2025
Viewed by 773
Abstract
Mercury, a prevalent heavy metal, negatively impacts oocyte maturation. However, the exact mechanism by which methylmercury chloride (MMC) affects this process remains elusive. The present study found that MMC administration triggered meiotic failure in oocytes by disrupting cumulus cell expansion, leading to compromised [...] Read more.
Mercury, a prevalent heavy metal, negatively impacts oocyte maturation. However, the exact mechanism by which methylmercury chloride (MMC) affects this process remains elusive. The present study found that MMC administration triggered meiotic failure in oocytes by disrupting cumulus cell expansion, leading to compromised spindle apparatus and altered chromosomal architecture, which are crucial for oocyte development. This disruption is characterized by abnormal microtubule organization and defective chromosome alignment. Additionally, MMC exposure caused oxidative stress-induced apoptosis due to mitochondrial dysfunction, as indicated by decreased mitochondrial membrane potential, mitochondrial content, mitochondrial DNA copy number, and adenosine triphosphate levels. Proteomic analysis identified 97 differentially expressed proteins, including P62, an autophagy marker. Our results confirmed that MMC induced autophagy, particularly through the hyperactivation of the mitochondrial autophagy to remove damaged and normal mitochondria. The mitochondrial reactive oxygen species (ROS) scavenger Mito-TEMPO alleviated oxidative stress and mitochondrial autophagy levels, suggesting that mitochondrial ROS initiates this autophagic response. Notably, MMC activates mitochondrial autophagy via the monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signal pathway due to mitochondrial dysfunction. In vivo studies in mice revealed that MMC exposure decreased reproductive performance, attributed to excessive mitochondrial autophagy leading to reduced oocyte quality. Overall, these findings demonstrate that MMC exposure impairs oocyte maturation via the hyperactivation of mitochondrial autophagy induced by mitochondrial dysfunction. Full article
(This article belongs to the Section Molecular Toxicology)
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14 pages, 3381 KiB  
Article
Recombinant Follicle-Stimulating Hormone and Luteinizing Hormone Enhance Mitochondrial Function and Metabolism in Aging Female Reproductive Cells
by Li-Te Lin, Chia-Jung Li, Yi-Shan Lee and Kuan-Hao Tsui
Int. J. Mol. Sci. 2025, 26(1), 83; https://doi.org/10.3390/ijms26010083 - 25 Dec 2024
Viewed by 1561
Abstract
Ovarian aging significantly impacts female fertility, with mitochondrial dysfunction emerging as a key factor. This study investigated the effects of recombinant follicle-stimulating hormone (FSH) and luteinizing hormone (LH) on mitochondrial function and metabolism in aging female reproductive cells. Human granulosa cells (HGL5) were [...] Read more.
Ovarian aging significantly impacts female fertility, with mitochondrial dysfunction emerging as a key factor. This study investigated the effects of recombinant follicle-stimulating hormone (FSH) and luteinizing hormone (LH) on mitochondrial function and metabolism in aging female reproductive cells. Human granulosa cells (HGL5) were treated with FSH/LH or not. Mitochondrial function was assessed through various assays, including mitochondrial mass, membrane potential, ROS levels, and ATP production. Mitochondrial dynamics and morphology were analyzed using MitoTracker staining. Cellular respiration was measured using a Seahorse Bioenergetics Analyzer. Metabolic reprogramming was evaluated through gene expression analysis and metabolite profiling. In vivo effects were studied using aging mouse oocytes. FSH/LH treatment significantly improved mitochondrial function in aging granulosa cells, increasing mitochondrial mass and membrane potential while reducing ROS levels. Mitochondrial dynamics showed a shift towards fusion and elongation. Cellular respiration, ATP production, and spare respiratory capacity were enhanced. FSH/LH-induced favorable alterations in cellular metabolism, favoring oxidative phosphorylation. In aging mouse oocytes, FSH/LH treatment improved in vitro maturation and mitochondrial health. In conclusion, FSH/LH supplementation ameliorates age-related mitochondrial dysfunction and improves cellular metabolism in aging female reproductive cells. Full article
(This article belongs to the Special Issue Reproductive Endocrinology Research)
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18 pages, 12347 KiB  
Article
In Vivo-Matured Oocyte Resists Post-Ovulatory Aging through the Hub Genes DDX18 and DNAJC7 in Pigs
by Cheng-Lin Zhan, Dongjie Zhou, Ming-Hong Sun, Wen-Jie Jiang, Song-Hee Lee, Xiao-Han Li, Qin-Yue Lu, Ji-Dam Kim, Gyu-Hyun Lee, Jae-Min Sim, Hak-Jae Chung, Eun-Seok Cho, Soo-Jin Sa and Xiang-Shun Cui
Antioxidants 2024, 13(7), 867; https://doi.org/10.3390/antiox13070867 - 19 Jul 2024
Viewed by 1863
Abstract
Assisted reproduction technology (ART) procedures are often impacted by post-ovulatory aging (POA), which can lead to reduced fertilization rates and impaired embryo development. This study used RNA sequencing analysis and experimental validation to study the similarities and differences between in vivo- and vitro-matured [...] Read more.
Assisted reproduction technology (ART) procedures are often impacted by post-ovulatory aging (POA), which can lead to reduced fertilization rates and impaired embryo development. This study used RNA sequencing analysis and experimental validation to study the similarities and differences between in vivo- and vitro-matured porcine oocytes before and after POA. Differentially expressed genes (DEGs) between fresh in vivo-matured oocyte (F_vivo) and aged in vivo-matured oocyte (A_vivo) and DEGs between fresh in vitro-matured oocyte (F_vitro) and aged in vitro-matured oocyte (A_vitro) were intersected to explore the co-effects of POA. It was found that “organelles”, especially “mitochondria”, were significantly enriched Gene Ontology (GO) terms. The expression of genes related to the “electron transport chain” and “cell redox homeostasis” pathways related to mitochondrial function significantly showed low expression patterns in both A_vivo and A_vitro groups. Weighted correlation network analysis was carried out to explore gene expression modules specific to A_vivo. Trait–module association analysis showed that the red modules were most associated with in vivo aging. There are 959 genes in the red module, mainly enriched in “RNA binding”, “mRNA metabolic process”, etc., as well as in GO terms, and “spliceosome” and “nucleotide excision repair” pathways. DNAJC7, IK, and DDX18 were at the hub of the gene regulatory network. Subsequently, the functions of DDX18 and DNAJC7 were verified by knocking down their expression at the germinal vesicle (GV) and Metaphase II (MII) stages, respectively. Knockdown at the GV stage caused cell cycle disorders and increase the rate of abnormal spindle. Knockdown at the MII stage resulted in the inefficiency of the antioxidant melatonin, increasing the level of intracellular oxidative stress, and in mitochondrial dysfunction. In summary, POA affects the organelle function of oocytes. A_vivo oocytes have some unique gene expression patterns. These genes may be potential anti-aging targets. This study provides a better understanding of the detailed mechanism of POA and potential strategies for improving the success rates of assisted reproductive technologies in pigs and other mammalian species. Full article
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22 pages, 4120 KiB  
Article
Three-Dimensionally Printed Agarose Micromold Supports Scaffold-Free Mouse Ex Vivo Follicle Growth, Ovulation, and Luteinization
by Emily J. Zaniker, Prianka H. Hashim, Samuel Gauthier, James A. Ankrum, Hannes Campo and Francesca E. Duncan
Bioengineering 2024, 11(7), 719; https://doi.org/10.3390/bioengineering11070719 - 15 Jul 2024
Cited by 4 | Viewed by 5394
Abstract
Ex vivo follicle growth is an essential tool, enabling interrogation of folliculogenesis, ovulation, and luteinization. Though significant advancements have been made, existing follicle culture strategies can be technically challenging and laborious. In this study, we advanced the field through development of a custom [...] Read more.
Ex vivo follicle growth is an essential tool, enabling interrogation of folliculogenesis, ovulation, and luteinization. Though significant advancements have been made, existing follicle culture strategies can be technically challenging and laborious. In this study, we advanced the field through development of a custom agarose micromold, which enables scaffold-free follicle culture. We established an accessible and economical manufacturing method using 3D printing and silicone molding that generates biocompatible hydrogel molds without the risk of cytotoxicity from leachates. Each mold supports simultaneous culture of multiple multilayer secondary follicles in a single focal plane, allowing for constant timelapse monitoring and automated analysis. Mouse follicles cultured using this novel system exhibit significantly improved growth and ovulation outcomes with comparable survival, oocyte maturation, and hormone production profiles as established three-dimensional encapsulated in vitro follicle growth (eIVFG) systems. Additionally, follicles recapitulated aspects of in vivo ovulation physiology with respect to their architecture and spatial polarization, which has not been observed in eIVFG systems. This system offers simplicity, scalability, integration with morphokinetic analyses of follicle growth and ovulation, and compatibility with existing microphysiological platforms. This culture strategy has implications for fundamental follicle biology, fertility preservation strategies, reproductive toxicology, and contraceptive drug discovery. Full article
(This article belongs to the Special Issue Bioengineering Technologies to Advance Reproductive Health)
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15 pages, 2400 KiB  
Article
The Simulated Physiological Oocyte Maturation (SPOM) System Enhances Cytoplasmic Maturation and Oocyte Competence in Cattle
by Micaela Navarro, Tomás Fanti, Nicolas Matias Ortega, Magalí Waremkraut, Francisco Guaimas, Adrian Ángel Mutto and Carolina Blüguermann
Animals 2024, 14(13), 1893; https://doi.org/10.3390/ani14131893 - 27 Jun 2024
Viewed by 1653
Abstract
In vitro embryo production is a widely applied technique that allows the expansion of genetics and accelerated breeding programs. However, in cattle, this technique still needs improvement in order to reach quality and pregnancy rates comparable to in vivo-derived embryos. One of the [...] Read more.
In vitro embryo production is a widely applied technique that allows the expansion of genetics and accelerated breeding programs. However, in cattle, this technique still needs improvement in order to reach quality and pregnancy rates comparable to in vivo-derived embryos. One of the limitations of this technique is related to in vitro maturation, where a heterogeneous population of oocytes is harvested from follicles and cultured in vitro in the presence of gonadotropic hormones to induce maturation. As a result, oocytes with different degrees of competence are obtained, resulting in a decrease in the quality and quantity of embryos obtained. A novel system based on the use of cyclic adenosine monophosphate (cAMP) modulators was developed to enhance bovine oocyte competence, although controversial results were obtained depending on the in vitro embryo production (IVP) system used in each laboratory. Thus, in the present work, we employed a reported cAMP protocol named Simulated Physiological Oocyte Maturation (SPOM) under our IVP system and analysed its effect on cytoplasmic maturation by measuring levels of stress-related genes and evaluating the activity and distribution of mitochondria as a marker for cytoplasmic maturation Moreover, we studied the effect of the cAMP treatment on nuclear maturation, cleavage, and blastocyst formation. Finally, we assessed the embryo quality by determining the hatching rates, total cell number per blastocyst, cryopreservation tolerance, and embryo implantation. We found that maturing oocytes in the presence of cAMP modulators did not affect nuclear maturation, although they changed the dynamic pattern of mitochondrial activity along maturation. Additionally, we found that oocytes subjected to cAMP modulators significantly improved blastocyst formation (15.5% vs. 22.2%, p < 0.05). Blastocysts derived from cAMP-treated oocytes did not improve cryopreservation tolerance but showed an increased hatching rate, a higher total cell number per blastocyst and, when transferred to hormonally synchronised recipients, produced pregnancies. These results reflect that the use of cAMP modulators during IVM results in competent oocytes that, after fertilisation, can develop in more blastocysts with a better quality than standard IVM conditions. Full article
(This article belongs to the Special Issue Livestock Reproduction: Reproductive Technologies in Animal Science)
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14 pages, 2387 KiB  
Article
Cytokine-Supplemented Maturation Medium Enhances Cytoplasmic and Nuclear Maturation in Bovine Oocytes
by Renata Blocher, Ying Liu, Tayler Patrick and Irina A. Polejaeva
Animals 2024, 14(12), 1837; https://doi.org/10.3390/ani14121837 - 20 Jun 2024
Cited by 2 | Viewed by 1956
Abstract
Bovine in vitro oocyte maturation (IVM) is an easy way to obtain oocytes for subsequent assisted reproductive techniques but is inefficient compared to in vivo maturation. Supplementation of three cytokines, fibroblast growth factor 2 (FGF2), leukemia inhibitory factor (LIF), and insulin-like growth factor [...] Read more.
Bovine in vitro oocyte maturation (IVM) is an easy way to obtain oocytes for subsequent assisted reproductive techniques but is inefficient compared to in vivo maturation. Supplementation of three cytokines, fibroblast growth factor 2 (FGF2), leukemia inhibitory factor (LIF), and insulin-like growth factor 1 (IGF1), or FLI, has increased oocyte maturation and embryo development in multiple species, but studies have not explored the oocyte differences caused by FLI IVM supplementation. This study aimed to assess important nuclear and cytoplasmic maturation events in high-quality oocytes. FLI-supplemented oocytes had a decreased GV (3.0% vs. 13.7%, p < 0.01) and increased telophase I incidence (34.6% vs. 17.6%, p < 0.05) after IVM, increased normal meiotic spindles (68.8% vs. 50.0%, p < 0.001), and an increased nuclear maturation rate (75.1% vs. 66.8%, p < 0.001). Moreover, in metaphase II oocytes, the percentage of FLI-treated oocytes with a diffuse mitochondrial distribution was higher (87.7% vs. 77.5%, p < 0.05) and with a cortical mitochondrial distribution was lower (11.6% vs. 17.4%, p < 0.05). Additionally, FLI-supplemented oocytes had more pattern I cortical granules (21.3% vs. 14.4%, p < 0.05). These data suggest that FLI supplementation in bovine in vitro maturation medium coordinates nuclear and cytoplasmic maturation to produce higher-quality oocytes. Full article
(This article belongs to the Special Issue Advances in In Vitro Oocyte Development in Domestic Animals)
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16 pages, 566 KiB  
Review
Allogenic Follicular Fosterage Technology: Problems, Progress and Potential
by Mingming Teng, Mengqi Zhao, Bo Mu and Anmin Lei
Vet. Sci. 2024, 11(6), 276; https://doi.org/10.3390/vetsci11060276 - 17 Jun 2024
Viewed by 1597
Abstract
The allogeneic follicular fosterage (AFF) technique transfers cumulus–oocyte complexes (COCs) from pubertal female animals to the dominant follicles of adult female animals for further development, allowing the COCs to further develop in a completely in vivo environment. This article reviews the history of [...] Read more.
The allogeneic follicular fosterage (AFF) technique transfers cumulus–oocyte complexes (COCs) from pubertal female animals to the dominant follicles of adult female animals for further development, allowing the COCs to further develop in a completely in vivo environment. This article reviews the history of AFF and JIVET and their effects on oocyte and embryo development as well as freezing resistance. Improving the efficiency and reproducibility of AFF technology is crucial to its clinical application. This article discusses factors that affect the success rate of AFF, including differences in specific technical procedures and differences between pubertal and adult follicles. Designing standardized procedures and details to improve the synchronization of donor COCs and recipient follicle maturity and reducing the damage to COCs caused by follicular aspiration may be the direction for improving the success rate of AFF in the future. Full article
(This article belongs to the Special Issue Advances in Veterinary Reproduction and Embryonic Development)
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15 pages, 3534 KiB  
Article
The Presence of TGFβ3 in Human Ovarian Intrafollicular Fluid and Its Involvement in Thromboxane Generation in Follicular Granulosa Cells through a Canonical TGFβRI, Smad2/3 Signaling Pathway and COX-2 Induction
by Tsung-Hsuan Lai, Hsuan-Ting Chen, Pi-Hui Wu and Wen-Bin Wu
Int. J. Mol. Sci. 2024, 25(10), 5558; https://doi.org/10.3390/ijms25105558 - 20 May 2024
Cited by 2 | Viewed by 1754
Abstract
Ovarian follicular fluid (FF) has a direct impact on oocyte quality, playing key roles in fertilization, implantation, and early embryo development. In our recent study, we found FF thromboxane (TX) to be a novel factor inversely correlated with oocyte maturation and identified thrombin, [...] Read more.
Ovarian follicular fluid (FF) has a direct impact on oocyte quality, playing key roles in fertilization, implantation, and early embryo development. In our recent study, we found FF thromboxane (TX) to be a novel factor inversely correlated with oocyte maturation and identified thrombin, transforming growth factor β (TGFβ), TNF-α, and follicular granulosa cells (GCs) as possible contributors to FF TX production. Therefore, this study sought to investigate the role of TGFβ3 in regulating TX generation in human ovarian follicular GCs. TGFβ3 was differentially and significantly present in the FF of large and small follicles obtained from IVF patients with average concentrations of 68.58 ± 12.38 and 112.55 ± 14.82 pg/mL, respectively, and its levels were correlated with oocyte maturity. In an in vitro study, TGFβ3 induced TX generation/secretion and the converting enzyme-COX-2 protein/mRNA expression both in human HO23 and primary cultured ovarian follicular GCs. While TGFβRI and Smad2/3 signaling was mainly required for COX-2 induction, ERK1/2 appeared to regulate TX secretion. The participation of Smad2/3 and COX-2 in TGFβ3-induced TX generation/secretion could be further supported by the observations that Smad2/3 phosphorylation and nuclear translocation and siRNA knockdown of COX-2 expression compromised TX secretion in GCs challenged with TGFβ3. Taken together, the results presented here first demonstrated that FF TGFβ3 levels differ significantly in IVF patients’ large preovulatory and small mid-antral follicles and are positively associated with oocyte maturation. TGFβ3 can provoke TX generation by induction of COX-2 mRNA/protein via a TGFβR-related canonical Smad2/3 signaling pathway, and TX secretion possibly by ERK1/2. These imply that TGFβ3 is one of the inducers for yielding FF TX in vivo, which may play a role in folliculogenesis and oocyte maturation. Full article
(This article belongs to the Special Issue Advances in Pharmacology of Prostaglandins)
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11 pages, 1683 KiB  
Article
Supplementation of SkQ1 Increases Mouse In Vitro Oocyte Maturation and Subsequent Embryonic Development by Reducing Oxidative Stress
by Zheng Li, Yiqian Zhang, Jinping Cao, Xupeng Xing, Yalin Liang, Yuxing Zhang, Xiaopeng Tang, Shengyi Lin, Zhenfang Wu, Zicong Li and Sixiu Huang
Pharmaceuticals 2024, 17(4), 455; https://doi.org/10.3390/ph17040455 - 2 Apr 2024
Cited by 5 | Viewed by 1912
Abstract
In vitro oocyte maturation (IVM) technology is important for assisted animal and human reproduction. However, the maturation rates and developmental potential of in vitro-matured oocytes are usually lower than those of in vivo-matured oocytes. Oxidative stress is a main factor that causes the [...] Read more.
In vitro oocyte maturation (IVM) technology is important for assisted animal and human reproduction. However, the maturation rates and developmental potential of in vitro-matured oocytes are usually lower than those of in vivo-matured oocytes. Oxidative stress is a main factor that causes the lower maturation rates and quality of in vitro-matured oocytes. The purpose of this study was to investigate the effects of treatment with SkQ1, a mitochondria-targeted antioxidant, on mouse IVM and subsequent embryonic development. The results demonstrated that the supplementation of SkQ1 during IVM improves the maturation rates of mouse oocytes and the subsequent developmental competence of in vitro-fertilized embryos. The addition of SkQ1 to the IVM medium also decreased oxidative stress and apoptosis, and increased mitochondrial membrane potential in matured mouse oocytes. This study provides a new method through which to enhance the maturation rates and the quality of in vitro-matured mouse oocytes, thus promoting the application and development of assisted animal and human reproductive technology. Full article
(This article belongs to the Section Pharmacology)
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