Theriogenology最新文献

{"title":"How can blood calcium concentration and oral supplementation affect uterine health in dairy cows?","authors":"Jerbeson Hoffmann da Silva , Clério Antônio Hoefle , Camila Cupper Vieira , Luiz Carlos Kreutz , André Gustavo Cabrera Dalto , Paulo Bayard Dias Gonçalves , Monique Tomazele Rovani , Rogério Ferreira","doi":"10.1016/j.theriogenology.2025.117496","DOIUrl":"10.1016/j.theriogenology.2025.117496","url":null,"abstract":"<div><div>A series of experiments was carried out to study the effects of blood calcium (Ca) concentration on the occurrence of subclinical endometritis (SE) in dairy cattle. In Experiment 1, serum Ca level of 26 cows in the first 24 h after calving was evaluated and correlated with the incidence of SE between 34 and 40 days postpartum. The polymorphonuclear leukocytes (PMN) count was affected by Ca blood concentration in the first 24 h postpartum (<em>P</em> ≤ 0.01). In Experiment 2, a total of 47 cows retrospectively classified as normocalcemic (Ca ≥8.5 mg/dL) or hypocalcemic (Ca <8.5 mg/dL) were randomly assigned to receive or not oral Ca formate at 6 and 30 h postpartum, and the effects of supplementation on SE incidence were evaluated. The treatment with Ca formate, regardless of Ca concentration, reduced PMN count and incidence of SE between 34 and 40 days postpartum (<em>P</em> < 0.05). For each increase of 1 mg/dL in blood Ca, SE incidence decreased by 22 %. In Experiment 3, five multiparous Holstein cows were enrolled in a 2x2 crossover model and assigned to 1 of 2 treatments: normocalcemic (Control) or induced subclinical hypocalcemia (iSCH) group. The sequence of treatments for a given cow was either Control-iSCH or iSCH-Control, followed by a 3-week washout period. The iSCH group received an intravenous infusion of 5 % disodium tetraacetic ethylenediamine Na2EDTA at 500 mL/h for 45 min, while the Control group received 0.9 % NaCl. Levels of ionized Ca (iCa), total Ca (tCa) and Mg were determined immediately before the treatments (M0), at 15 (M1), 45 min (M2, end of infusions) and 3 h after the end of treatments (M3). Furthermore, glucose, insulin, NEFA and BHB levels were determined at M3. Immediately after the end of the infusions (M2), all animals received an intrauterine challenge with 300 μg of LPS and, after 3 h (M3), an endometrial biopsy was performed to determine the expression of <em>IL-6</em>, <em>CXCL8</em> and <em>TNF</em> and gene expression of glucose-insulin receptors and insulin-related genes. Subclinical hypocalcemia was effectively induced; however, the expression of <em>IL-6</em>, <em>CXCL8</em> and <em>TNF</em> in the endometrium did not differ between groups within 3 h after intrauterine LPS challenge. The iSCH group presented a significant reduction in serum BHB levels compared to the Control group, but there were no differences in serum glucose, insulin, NEFA and in the endometrial expression of the <em>INSR, IRS1, IGF1, SLC2A1</em>, and <em>SLC2A3</em> genes. In conclusion, PMN infiltration and the incidence of SE from 34 to 40 days postpartum is related to blood Ca immediately after calving. Ca supplementation with Ca formate decreases the number of PMNs in the endometrium, reducing SE incidence. In addition, transient iSCH does not cause immediate significant changes in the endometrial expression of key inflammatory cytokines and does not adversely affect the insulin signaling pathway in the uterus with","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"244 ","pages":"Article 117496"},"PeriodicalIF":2.4,"publicationDate":"2025-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144115673","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Single-dose recombinant FSH as a viable alternative to multi-dose porcine FSH in commercial superovulation protocols in cows","authors":"Monique Mazzarollo Frata ,&nbsp;Wagner Marques de Lima ,&nbsp;Monique Tomazele Rovani ,&nbsp;Rogério Ferreira ,&nbsp;Alexandre Hênryli de Souza ,&nbsp;Rafael Gianella Mondadori ,&nbsp;Paulo Bayard Dias Gonçalves ,&nbsp;Vilceu Bordignon ,&nbsp;Bernardo Garziera Gasperin","doi":"10.1016/j.theriogenology.2025.117498","DOIUrl":"10.1016/j.theriogenology.2025.117498","url":null,"abstract":"<div><div>Superovulation (SOV) for embryo transfer (ET) typically requires eight decreasing doses of porcine FSH (pFSH) due to the short half-life. Recently, a recombinant FSH (rFSH) formulation has become available, allowing for SOV with a single treatment. This study aimed to compare the outcomes of SOV in commercial ET programs using eight doses of pFSH versus a single administration of rFSH. Data from 569 SOV procedures conducted under commercial conditions with pFSH (n = 291) or rFSH (n = 278) were evaluated. No significant differences were observed regarding the total number of recovered structures (TS), degenerated embryos (DG), and transferable embryos (TE). However, the number and proportion of unfertilized oocytes (UF) relative to TS were higher in the rFSH group. Conversely, the proportion of DG relative to TS was higher in the pFSH group. The proportion of TE relative to TS was consistent at 60 % for both treatments. Finally, when a small subset of cows (n = 12) were SOV four times with rFSH at 40- to 60-day intervals, no significant differences were observed in TS or TE. In conclusion, a single administration of rFSH provides SOV outcomes comparable to those achieved with eight administrations of pFSH in commercial ET programs and results seem constant across repeated treatments with rFSH.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"244 ","pages":"Article 117498"},"PeriodicalIF":2.4,"publicationDate":"2025-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144098417","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Gonadotropin inhibitory hormone (GnIH) induces glucose metabolism disorders and ovarian dysfunction in ovarian white follicles of hens","authors":"Bingqian Shen ,&nbsp;Dongyang Han ,&nbsp;Yuanyuan Xin ,&nbsp;Jiani Zhang ,&nbsp;Changlin Xu ,&nbsp;Xingxing Song ,&nbsp;Xin Zhang ,&nbsp;Chengcheng Liu ,&nbsp;Jiang Li ,&nbsp;Yixian Wei ,&nbsp;Xiaoye Wang ,&nbsp;Chuanhuo Hu ,&nbsp;Xun Li","doi":"10.1016/j.theriogenology.2025.117497","DOIUrl":"10.1016/j.theriogenology.2025.117497","url":null,"abstract":"<div><div>Energy metabolism is crucial for reproduction, and disturbances in glucose metabolism are closely associated with reproductive disorders. The gonadotropin inhibitory hormone (GnIH) plays a crucial role in reproduction and glucose homeostasis in both birds and mammals. However, its specific effects on glucose metabolism-associated ovarian dysfunction in hens remain uncharacterized. In this study, we investigated the effects of GnIH on ovarian function and glucose homeostasis in hens using combined <em>in vivo</em> and <em>in vitro</em> approaches. Our results showed that GnIH and its receptor, GPR147, are predominantly expressed in white follicles. Continuous GnIH injection significantly reduced the ovarian index and the number of prehierarchical follicles, concurrently suppressing Fshr and LHr expression in white follicles. Additionally, metabolomic analyses indicate changes in key glucose metabolism pathways, indicating a regulatory role of GnIH in glucose metabolism. To validate our metabolomics findings, the effects of GnIH on glucose homeostasis in ovarian white follicles and granulosa cells (GCs) were further investigated through <em>in vivo</em> and <em>in vitro</em> studies. The results demonstrated that GnIH enhanced glucose transport, glycolysis and glycogen synthesis, but concurrently induced insulin resistance, oxidative stress, and mitochondrial dysfunction, ultimately leading to reduced energy levels and apoptosis in ovarian tissues. In summary, our study reveals that GnIH contributes to ovarian dysfunction via glucose metabolism dysregulation in hens, suggesting its potential as a therapeutic target for metabolic disorder-associated ovarian impairment in chickens.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"244 ","pages":"Article 117497"},"PeriodicalIF":2.4,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144115671","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Canine Brachycephalic Obstructive Airway Syndrome impacts respiratory and hemodynamic physiological adaptations during pregnancy","authors":"Thayná Pantoja Gardés,&nbsp;Roberto Rodrigues da Rosa Filho,&nbsp;Maíra Morales Brito,&nbsp;Leticia Lima de Almeida,&nbsp;Renato Bueno Flores,&nbsp;Renata Azevedo de Abreu,&nbsp;Camila Infantosi Vannucchi","doi":"10.1016/j.theriogenology.2025.117495","DOIUrl":"10.1016/j.theriogenology.2025.117495","url":null,"abstract":"<div><div>Brachycephalic obstructive airway syndrome (BOAS) has a negative impact on animal welfare and quality of life, with perturbing signs of respiratory distress and both clinical and reproductive issues. Therefore, the aim of this study was to compare the respiratory pattern, uterine hemodynamic profile, and placenta development during different stages of pregnancy in bitches with or without symptoms of the brachycephalic obstructive airway syndrome. Bitches were grouped according to the symptomatic manifestation of BOAS before breeding, as Brachycephalic Syndrome Group (n = 10) and Non-brachycephalic Syndrome (Control) Group (n = 10). Along gestation (early, mid and late stage), females were examined for clinical evaluations (heart and respiratory rates, body temperature, capillary refill time, hydration and mucosal color score, blood pressure and peripheral oxygen saturation), arterial blood acid-base balance and obstetric B-mode and uterine and umbilical artery dopplervelocimetry. BOAS bitches had higher (P &lt; 0.05) early-pregnancy heart rate, and increased (P &lt; 0.05) respiratory rate at late-pregnancy, tendency to acidemia and lower base excess (BE) and bicarbonate concentration until mid-pregnancy, while PCO₂ remained unchanged. Fetus of BOAS females had slower (P &lt; 0.05) placental growth and improvement of umbilical artery blood flow along gestation, although uterine artery vascularization remained unchanged. In conclusion, pregnant BOAS bitches have altered respiratory pattern and acid-base balance during pregnancy; however, without negatively impacting hemodynamics of the umbilical artery. Adaptations within the fetal-placental unit were sufficient to meet physiological pregnancy demands and partially support proper fetal growth.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"244 ","pages":"Article 117495"},"PeriodicalIF":2.4,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144115672","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effects of polystyrene nanoparticles on bovine oocyte in vitro maturation","authors":"Merlo Barbara ,&nbsp;Alessandro Marino Volsa ,&nbsp;Laura Tovar ,&nbsp;Margherita Gaiani ,&nbsp;Penelope Maria Gugole ,&nbsp;Emilia Attolini ,&nbsp;Iacono Eleonora","doi":"10.1016/j.theriogenology.2025.117482","DOIUrl":"10.1016/j.theriogenology.2025.117482","url":null,"abstract":"<div><div>Polystyrene nanoparticles (PS-NPs) have emerged as a significant environmental concern due to their widespread presence and potential toxicity. This study investigates the effects of PS-NPs on bovine oocyte in vitro maturation (IVM) to assess their impact on reproductive health. During IVM, oocytes were exposed to varying concentrations of 100 nm PS-NPs (0, 5, 50, 100, and 200 μg/mL) or of fluorescent 30 nm PS-NPs (0, 5, 35, and 70 μg/mL). Higher PS-NPs concentrations (100 and 200 μg/mL for 100 nm PS-NPs and 70 μg/mL for 30 nm PS-NPs) significantly reduced nuclear maturation rates and increased degeneration. Similarly, cytoplasmic maturation was negatively affected at higher concentrations (P &lt; 0.05). PS-NPs exposure also elevated reactive oxygen species (ROS) levels and reduced glutathione (GSH) content within the oocyte. Notably, PS-NPs were internalized by both oocytes and cumulus cells in a concentration-dependent manner. These findings confirm the reproductive toxicity of PS-NPs, emphasizing their potential to compromise mammalian fertility and raising concerns regarding their environmental and health implications.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"244 ","pages":"Article 117482"},"PeriodicalIF":2.4,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144069173","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sperm fatty acid composition as an indicator of quality and cryotolerance in blue catfish, Ictalurus furcatus","authors":"Kyle R. Wood ,&nbsp;Matthew K. Litvak ,&nbsp;Samitha S.N. Liyanage ,&nbsp;Kaylan A. Martin ,&nbsp;V. MacKenzie Tackett ,&nbsp;Rex A. Dunham ,&nbsp;Luke A. Roy ,&nbsp;Cynthia K. Faulk ,&nbsp;Benjamin H. Beck ,&nbsp;Jason W. Abernathy ,&nbsp;Xu Wang ,&nbsp;Ian A.E. Butts","doi":"10.1016/j.theriogenology.2025.117487","DOIUrl":"10.1016/j.theriogenology.2025.117487","url":null,"abstract":"<div><div>There is a lack of biomarkers that can be used to predict fresh and frozen-thawed sperm quality for the commercially important blue catfish, <em>Ictalurus furcatus.</em> Sperm membranes are composed of various fatty acids (FAs) that have been linked to reproductive success across other animal and fish taxa, making it a potential biomarker. The objectives of this study were to (i) compare sperm FA percentages between fresh and cryopreserved male groups; and (ii) use sperm kinematics of fresh and cryopreserved males to create “good” (i.e. fast swimming sperm kinematics) and “bad” (i.e. slow swimming sperm kinematics) quality groups, then compare FA percentages of the groups to pinpoint physiological biomarkers of fresh and frozen-thawed sperm quality and cryotolerance. We found significant differences between fresh and cryopreserved sperm where fresh sperm had significantly higher ratios of n-3:n-6 and EPA:ARA as well as n-3 PUFAs and MUFAs, while cryopreserved sperm had higher concentrations of saturates. Cryopreserved sperm from good and bad males also had differences, such as a significantly higher concentration of saturates in good males and significantly higher concentrations of MUFAs, n-6 PUFAs, n-3 PUFAs, and total PUFAs in bad males. Lastly, low levels of MUFAs and n-6 PUFAs in fresh sperm resulted in higher post-thaw sperm kinematics. This knowledge can now be used to create additional biomarkers of sperm quality and to formulate the FA profile of catfish diets to improve sperm quality, thereby, improving their ability to handle the stressors of the cryopreservation process.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"245 ","pages":"Article 117487"},"PeriodicalIF":2.4,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144134073","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Furosemide sodium enhanced AQP3 expression by inhibiting NKCC1 leading to improved vitrification freezing rates of mouse and bovine blastocysts","authors":"Chang-Guo Min ,&nbsp;Xue-Song Shan ,&nbsp;Kai-Yan Zhang ,&nbsp;Yu-Chan Wang ,&nbsp;Xin Ma ,&nbsp;Jun Wang ,&nbsp;Jing Zhao ,&nbsp;Yi Fang ,&nbsp;Hong-Yu Liu ,&nbsp;He Ding ,&nbsp;Dian Wang ,&nbsp;Jing Guo ,&nbsp;Wen-Fa Lu","doi":"10.1016/j.theriogenology.2025.117485","DOIUrl":"10.1016/j.theriogenology.2025.117485","url":null,"abstract":"<div><div>Furosemide sodium, a loop diuretic that inhibits the Na-K-2Cl cotransporter (NKCC1), is commonly used in clinical treatment of edema. In this study, we investigated whether furosemide sodium could improve the survival and quality of vitrified mouse and bovine blastocysts by promoting the expulsion of blastocoelic fluid, thus enhancing cryoprotectant penetration and reducing ice crystal formation. Mouse and bovine blastocysts were vitrified using equilibration solutions (ES) and vitrification solutions (VS) with or without furosemide sodium, then thawed for survival and quality assessment. Results indicated that furosemide sodium significantly improved post-thaw survival rates (<em>P</em> &lt; 0.05). It also markedly reduced reactive oxygen species (ROS) levels (<em>P</em> &lt; 0.05). Additionally, furosemide sodium increased glutathione (GSH) levels (<em>P</em> &lt; 0.05) and enhanced mitochondrial function. The improvement in mitochondrial function was reflected in increased mitochondrial membrane potential, which are key indicators of mitochondrial health and energy status (<em>P</em> &lt; 0.05). Immunofluorescence analysis revealed that furosemide sodium downregulated NKCC1 protein levels and upregulated Aquaporin 3 (AQP3) protein levels (<em>P</em> &lt; 0.05), indicating that furosemide sodium modulates ion transport and water regulation. This modulation likely contributes to the closure of sodium-potassium ion channels, reducing ion influx and promoting water efflux from the blastocyst. This closure limited extracellular sodium and potassium influx, significantly reducing these ions within the blastocyst cavity (<em>P</em> &lt; 0.05). Similar effects were observed in both mouse and bovine blastocysts, where furosemide sodium promoted fluid expulsion from the blastocoel, thereby enhancing vitrification efficiency. These findings suggested that furosemide sodium may be a promising agent to improve vitrification efficiency in blastocysts by increasing osmotic pressure differences across the blastocoel.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"244 ","pages":"Article 117485"},"PeriodicalIF":2.4,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144070257","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Interleukin-6 supplementation during in vitro maturation improves developmental competence of in vitro fertilized porcine oocytes","authors":"Manuela Garcia-Canovas ,&nbsp;Adelina Lopez-Jara ,&nbsp;Alejandro Gonzalez-Plaza ,&nbsp;Inmaculada Parrilla ,&nbsp;Heriberto Rodriguez-Martinez ,&nbsp;Cristina Cuello ,&nbsp;Emilio A. Martinez ,&nbsp;Maria A. Gil","doi":"10.1016/j.theriogenology.2025.117483","DOIUrl":"10.1016/j.theriogenology.2025.117483","url":null,"abstract":"<div><div>Interleukin 6 (IL-6) is a cytokine physiologically expressed within the reproductive tract, where it plays a crucial role in cellular communication and processes such as ovulation and implantation. These processes and the presence of embryos alter the immune response of the female reproductive tract in vivo, a physiological event of utmost relevance. Mimicking such physiological environment may optimize porcine in vitro embryo production (IVP). This study aimed to assess the impact of IL-6 supplementation during in vitro maturation (IVM) and/or in vitro culture (IVC) on the developmental competence of pig oocytes fertilized in vitro. In the first experiment, cumulus-oocytes complexes (COCs) were matured in media containing graded IL-6 concentrations (0–200 ng/mL). IL-6 did not affect maturation, fertilization, cleavage, or blastocyst formation rates. However, 100 ng/mL IL-6 significantly (<em>P</em> &lt; 0.05) improved chromosome alignment, meiotic spindle formation, blastocyst efficiency, and total cell number compared to controls. In the second experiment, COCs were matured and fertilized without IL-6 supplementation. The resulting presumptive zygotes were cultured in IVC media with graded concentrations of IL-6 (0–200 ng/mL). IL-6 supplementation during IVC did not enhance developmental competence. Conversely, the highest IL-6 concentration significantly (<em>P</em> &lt; 0.05) reduced cleavage rate. In the third experiment, combined supplementation of 100 ng/mL IL-6 during IVM and 10 ng/mL during IVC showed no additional improvements compared to IL-6 supplementation during IVM alone. In conclusion, the results of this study indicate that IL-6, particularly when added during oocyte maturation, positively impacts porcine in vitro embryo production.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"244 ","pages":"Article 117483"},"PeriodicalIF":2.4,"publicationDate":"2025-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144069171","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Induction of feline fetal fibroblasts into pluripotent stem cells using cat-derived reprogramming factors","authors":"Jian Qin ,&nbsp;Nannan Sun ,&nbsp;Yitong Wang ,&nbsp;Jie An ,&nbsp;Dipeng Zhao ,&nbsp;Junling Li ,&nbsp;Hao Zhang ,&nbsp;Rong Du","doi":"10.1016/j.theriogenology.2025.117481","DOIUrl":"10.1016/j.theriogenology.2025.117481","url":null,"abstract":"<div><div>There are few studies on the establishment of induced pluripotent stem cells (iPSCs) in cats. Although induction using heterologous reprogramming factors delivered via viral vectors has been reported, its safety and reprogramming efficiency still require improvement. In addition, the reprogramming mechanism needs further elucidation. In this study, we constructed a series of expression vectors for cat-derived reprogramming transcription factors based on the piggyBac transposon system and transfected various factor combinations into cat fetal fibroblasts (CFFs) under different electroporation conditions to generate cat iPSCs (ciPSCs). Additionally, the specific roles of these factors in reprogramming were investigated. The results showed that under the optimized electroporation conditions (DMEM/F12 buffer, 300 V, 10 ms pulse duration, 2 pulses, 25 μg plasmid DNA, and 4 mm cuvette), the survival rate and transfection efficiency of CFFs reached 64 % and 67.8 %, respectively. Based on this condition, a seven-factor combination (cOSKM + pNL + SV40 Large T) was confirmed as a better inducer for establishing ciPSCs. The obtained ciPSCs exhibit good pluripotency and passaging stability. They express stemness-related genes and proteins, and can form embryoid bodies (EBs) capable of differentiating into all three germ layers. OCT4 (O), SOX2 (S), KLF4 (K), and c-MYC (M) play important cooperative and synergistic roles in the mesenchymal-to-epithelial transition (MET) during the initial stages of reprogramming, while the supplement of NANOG (N) and LIN28 (L) can further promote MET and is important for successful reprogramming. It lays a foundation for the further breeding of cloned and genetically modified cats, and provides a tool for studying embryonic developmental diseases, screening drugs, and applying to tissue regeneration.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"244 ","pages":"Article 117481"},"PeriodicalIF":2.4,"publicationDate":"2025-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144070256","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cryoprotective effects of mesenchymal stem cell and seminal plasma-derived extracellular vesicles on canine sperm","authors":"Merve Deniz Tanrıkulu ,&nbsp;Mesut Çevi̇k ,&nbsp;Melek Yüce ,&nbsp;Pakize Neslihan Taşlı ,&nbsp;Kubilay Yıldırım","doi":"10.1016/j.theriogenology.2025.117480","DOIUrl":"10.1016/j.theriogenology.2025.117480","url":null,"abstract":"<div><div>In this study, the lack of standardized freezing protocols for sperm cryopreservation in dogs and the limited research on using exosomes in cryopreservation were considered. Additionally, unlike previous studies on sperm cryopreservation, we introduced an innovative approach using the Aqueous Two-Phase System (ATPS) method for exosome isolation. This study aimed to evaluate the sperm-protective effects of adipose tissue-derived mesenchymal stem cell exosomes (MSC-exo) and seminal plasma exosomes (SP-exo) in dog sperm cryopreservation. Ejaculates from six dogs were processed with Tris-based diluents and divided into four groups: MSC-exo, 1.5 % SP-exo, 2 % SP-exo, and control, and frozen. After thawing, sperm motility, viability, membrane integrity, chromatin integrity, morphological integrity, and gene expression levels were analyzed. The results showed that the MSC-exo group had significantly higher total motility (%60.31 ± 6.12), progressive motility (%22.09 ± 3.34), plasma membrane integrity (%66.94 ± 2.24), and viability (%70.88 ± 1.95) compared to the other groups (P &lt; 0.05). Additionally, the normal chromatin packaging rate was highest in the MSC-exo group (%91.33 ± 0.61, P &lt; 0.05). While some improvements were observed in the SP-exo groups, they were not as pronounced as in the MSC-exo group. No significant differences were found in gene expression levels, although an improvement trend was observed in the MSC-exo group. In conclusion, MSC-exo reduced cryopreservation-induced sperm damage and provided overall protection in sperm parameters. These findings suggest that MSC-exo could be a potential biological additive in dog sperm freezing protocols.</div></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":"244 ","pages":"Article 117480"},"PeriodicalIF":2.4,"publicationDate":"2025-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144069172","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}