Romero Aguirregomezcorta, Jon

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Romero Aguirregomezcorta, Jon

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Romero Aguirregomezcorta, Jon

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Anatomía y Anatomía Patológica Comparadas

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Open Access
Efecto de las secreciones de organoides de endometrio humano sobre el cultivo in vitro de embriones bovinos
(Facultad de Veterinaria y el Servicio de Publicaciones de la Universidad de Murcia, 2023) Masegosa Domínguez, Alicia María; Navarro-Serna, Sergio; Pérez-García, Vicente; Coy, Pilar; Juan, Ángela; Romero Aguirregomezcorta, Jon
Durante los últimos 50 años, la fertilidad de las vacas lecheras de alta producción ha disminuido debido a la intensidad de la selección genética. Para superar este obstáculo, se ha recurrido a las técnicas de reproducción asistida. Sin embargo, la producción de embriones in vitro es un proceso relativamente ineficiente, con tasas máximas de blastocistos obtenidos de alrededor del 35%. Para solventar este problema, se planteó la hipótesis de añadir al medio de cultivo secreciones de organoides de endometrio, así, los embriones producidos in vitroestarían expuestos a un ambiente más fisiológico y el rendimiento de la técnica aumentaría. Para ello, se llevó a cabo la recolección de ovocitos bovinos procedentes de un matadero local, y se procedió a su maduración in vitro, fecundación in vitro y cultivo embrionario in vitro. El medio de cultivo embrionario se suplementó al 1% (v/v) con i) secreciones de organoides de endometrio humano (ya que no existe disponibilidad de secreciones bovinas), ii) medio de cultivo Boretto, y ii) gotas de matrigel con medio de cultivo Boretto, como controles de procedimiento. Además, se incluyó un grupo control sin ningún suplemento. Se evaluó el desarrollo embriona-rio a los días 2, 7 y 8 de cultivo. El porcentaje de blastocistos a día 7 y a día 8 post inseminación fue significa-tivamente mayor (P<0.05) cuando se añadió el medio de cultivo Boretto. Además, este tratamiento incrementó la cinética de desarrollo de los embriones a día 7 post inseminación, y la calidad de los blastocistos evaluada por el número total de células, con un 85.8% más que el control. Aunque las secreciones de endometrio humano no han dado el resultado esperado, sería necesario comprobar si las secreciones de organoides específicas de endometrio bovino, a distintas concentraciones, podrían incrementar el rendimiento de la producción in vitro de embriones bovinos.
Open Access
Long-term haematological analysis in pigs derived from assisted reproductive technologies
(Brazilian College of Animal Reproduction, 2025) Mikita, Małgorzata; Cánovas, Sebastián; Heras, Sonia; Ramis, Guillermo; Gadea, Joaquín; Coy, Pilar; Romar, Raquel; Romero Aguirregomezcorta, Jon; Anatomía y Anatomía Patológica Comparadas
Rising livestock demand has increased use of assisted reproductive technologies (ART). Evidence indicates long-term effects of ART on molecular physiology and metabolism. This study explores ART’s impact on haematological parameters in a colony of pigs from 1 to 5 years of age. Animals were born after artificial insemination (AI) and transfer of embryos in vitro produced with (RF-IVP) or without (C-IVP) oviductal and uterine fluids during in vitro fertilization and embryo culture media (París-Oller et al., J Anim Sci and Biotechnol, 2021). Pigs were maintained under identical conditions and sampled every six months. Number of animals in each group decreased throughout life: 33 to 9 (AI); 22 to 10 (C-IVP); and 14 to 4 (RF-IVP). The number of blood samples was the same as the number of animals and were collected via jugular venipuncture in lithium heparin tubes and immediately transported to laboratory. A haematology analyzer (Siemens ADVIA® 120, USA) assessed red blood cell (RBC) count, haemoglobin concentration (HB), haematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), cell haemoglobin concentration mean (CHCM), red blood cell distribution width (RDW), haemoglobin concentration distribution width (HDW), white blood cell (WBC) count; and the differential count of neutrophils, lymphocytes, monocytes, eosinophils, and basophils. Reticulocyte percentage, mean corpuscular volume of reticulocytes (MCVr), haemoglobin content in reticulocytes (CHr) and platelet count were analyzed. Data were analyzed by mixed-effects model and likelihood-ratio test to determine effects of age, group and sex on the variables. Post-hoc test for multiple comparisons Tukey’s method were used. P<0.05 was considered significant. Age affected all variables. MCH, CH, CHCM, WBC, and platelet indices (MPV, MPC, MPM, PMDW) increased through life. Biphasic trajectories occurred in MCV, HTC, RBC, neutrophil counts, monocyte counts, PLT counts, and lymphocytes (secondyear decline followed by increase). Progressive decline with age was observed in monocyte percentage and reticulocyte parameters. Irregular oscillatory patterns dominated HB, RDW, MCHC, CHDW, HDW, eosinophil and basophil parameters, and advanced platelet indices (large PLT, PCT, PDW, MCVr, PCDW). HCT, RBC, and HB reached lowest values in AI group, highest in RF-IVP. HCM, MCHC, and lymphocyte percentage showed lowest means in AI, highest in C-IVP. RDW, WBC, neutrophil parameters, and lymphocyte count were lowest in C-IVP, highest in AI. Monocyte count exhibited lowest values in C-IVP, highest in RF-IVP group. CH and PCDW were higher in females than males, whereas HDW, PDW, MPC were higher in males. Long-term results confirm slight persisting haematological differences in naturally and artificially conceived pigs, although the clinical relevance is unnoticeable.
Open Access
Deciphering differences in DNA methylation and transcriptome profiles of oocytes from pigs with high and low developmental competence
(Oxford University Press, 2025-06-03) Abril Parreño, Laura; Lopes, Jordana S.; Romero Aguirregomezcorta, Jon; Galvao, Antonio; Kelsey, Gavin; Coy Fuster, Pilar; Fisiología
In vitro maturation (IVM) is a critical step in animal in vitro embryo production, yet oocytes matured in vitro often exhibit lower developmental competence than their in vivo counterparts. However, the molecular mechanisms behind this observation remain unclear. This study investigated the gene expression and DNA methylation profiles in porcine oocytes with different developmental competencies. To study these differences, we used as model oocytes from prepubertal gilts (IVM) and sows (in vivo matured) and assessed their developmental competence up to the blastocyst stage. We also examined their gene expression and DNA methylation profiles at single-cell resolution using RNA-sequencing and bisulfite-sequencing. Oocytes were obtained by aspiration of either ovarian follicles between 3-6mm diameter, and the subsequent IVM, or ovarian follicles from 8-10 mm diameter, with no need for maturation (in vivo matured oocytes). Cleavage rates (58.2±3.0 and 45.7±4.4) and blastocyst rates (31.4±3.7 and 47.5±6.6) for IVM and in vivo groups differed significantly. Using the in vivo group as a reference, IVM oocytes had 1297 down-regulated and 476 up-regulated differentially expressed genes (DEGs), with up-regulated DEGs associated with organelle organization and cell cycle processes, and down-regulated genes involved in protein synthesis and metabolomic processes. While global DNA methylation levels were similar between groups, a few differentially methylated regions were found in CpG islands, promoters, and coding regions. Our integrative analysis identified key methylated regions and genes that distinguish each group, suggesting that both donor age and maturation conditions significantly influence gene expression regulation in oocytes with different developmental competencies.
Open Access
Metabolic profile and glycemic response in fully-grown sows born using assisted reproductive technologies
(Elsevier, 2024-10-02) Cánovas Bernabé, Sebastián; Heras García, Sonia; Romero Aguirregomezcorta, Jon; Quintero Moreno, Armando Arturo; Gadea Mateos, Joaquín; Coy Fuster, Pilar; Romar Andrés, Raquel; Anatomía y Anatomía Patológica Comparada; Facultad de Veterinaria
The aim of the present work was to gain insight into the metabolism of pigs derived from assisted reproductive technologies during their adulthood. Approximately 4h after feeding, a blood sample was taken from 3.5 year old sows born by artificial insemination (AI group, n = 7) and transfer of in vitro produced embryos (IVP group, n = 11) to determine the physiological concentrations of the main biomarkers of carbohydrates (glucose and lactate), proteins (albumin, creatinine and urea) and lipids (cholesterol and triglycerides). Four weeks later, an oral glucose tolerance test (OGTT; 1.75g glucose/kg body weight) was performed after an overnight fast and 1h of water withdrawal. Blood samples were obtained prior (T = 0 min; fasting conditions) and 15, 30, 45, 60, 90, 120, 150, 180, 210 and 240 min after glucose intake. At each time point, glycemia was measured immediately using glucometer test strips, and serum was collected to determine the above metabolites along with insulin and glucagon. After OGTT, the area under the curve (AUC) between sampling times and homeostasis model assessment of insulin resistance (HOMA) indices were calculated. Under physiological conditions, the concentration of metabolites studied was similar between AI and IVP sows. In both groups, fasting decreased cholesterol and increased triglycerides and urea (P < 0.001). However, creatinine and lactate were similar in both groups under physiological and fasting conditions. The expected increase in albuminemia and decrease in glycaemia after fasting was only observed in IVP sows. OGTT revealed a different glucose curve pattern (monophasic in AI and biphasic in IVP group), a lower mean concentration of cholesterol, glucose, lactate, triglycerides in IVP compared to AI pigs (P < 0.01), and a higher mean concentration of albumin, creatinine and insulin in IVP compared to AI group (P < 0.05). On the contrary, no differences were found between groups for mean serum glucagon and urea levels, nor for glucose homeostasis indices HOMA-IR and HOMA-%B. The AUC differed between groups at several time points with larger AUC for creatinine, and smaller AUC for glucose, glucagon, and triglycerides, in IVP pigs than in AI pigs at 180–210 min (P < 0.05). In conclusion, under physiological conditions the metabolic profile of fully-grown AI and IVP sows is similar and within normal ranges. Glucose challenge revealed differences in metabolic and insulin responses between groups but with normal glucose tolerance in both cases.
Open Access
Respuesta a la estimulación ovárica mediante FSH (folltropin®) y rendimiento de opu en vacas adultas obtenidas por diferentes técnicas de reproducción asistida
(Universidad de Murcia. Servicio de publicaciones, 2022-11-15) Morera Jiménez, Alba; Velasco García, Elena; Ruiz López, Salvador; Romero Aguirregomezcorta, Jon; Anatomía y Anatomía Patológica Comparadas
Los objetivos de este trabajo han sido realizar un seguimiento ecográfico de los ovarios bovinos para comprobar la respuesta folicular de vacas adultas nacidas por diferentes técnicas de reproducción asistida (TRA), tras aplicar un protocolo de estimulación ovárica con FSH (Folltropin®); y determinar la eficacia reproductiva de los animales estudiados, analizando la cantidad y calidad de los ovocitos obtenidos por aspiración folicular (Ovum Pick-Up, OPU) antes y después del protocolo de estimulación empleado. La experiencia se ha llevado a cabo en las instalaciones del santuario bovino de la Granja Docente Veterinaria de la Universidad de Murcia. Se han utilizado 4 hembras bovinas obtenidas por distintas TRA: inseminación artificial (IA), transferencia de embriones (TE) producidos por TRA con ovocitos cultivados in vitro en medios convencionales y por TE obtenidos por TRA con ovocitos cultivados con fluidos naturales (folicular y oviductal).Los animales fueron estimulados durante 5 semanas mediante un protocolo que consistía en la aplicación de GnRH y posteriormente, FSH (Folltropin®, 350 UI/animal) durante 3 días en dosis decrecientes. Una vez a la semana, al finalizar el protocolo de estimulación ovárica, se realizaba una ecografía de los ovarios y una aspiración folicular ecoguiada (OPU) para comprobar la respuesta al protocolo de estimulación y determinar el número de folículos y de ovocitos obtenidos tras la OPU y su calidad ovocitaria. Previamente al periodo de estimulación con FSH, las vacas fueron ecografiadas y sometidas a OPU 3 semanas sin estimulación ovárica como grupo control.A lo largo del trabajo realizado, se ha comprobado la respuesta al protocolo de estimulación ovárica ensa-yado, y se ha determinado la eficacia reproductiva de los animales, según el número de folículos aspirados y la cantidad y calidad de los ovocitos obtenidos por OPU en cada una de ellas.
Open Access
Respuesta a la estimulación ovárica mediante FSH (Folltropin®) y rendimiento de OPU en vacas adultas obtenidas por diferentes técnicas de reproducción asistida
Morera-Jiménez, Alba; Velasco-García, Elena; Heras, Sonia; Ruiz, Salvador; Romero Aguirregomezcorta, Jon; Dpto. Fisiología. Facultad de Veterinaria, Complejo Espinardo. 30100. Universidad de Murcia. Murcia. España.
Open Access
Long term study of the blood plasma biochemical profile of cattle born by assisted reproductive technologies
(Brazilian College of Animal Reproduction, 2024) Serrano-Albal, Maria; Canovas, Sebastian; Heras, Sonia; Quintero-Moreno, Armando Arturo; Lopes, Jordana Sena; Soriano-Ubeda, Cristina; Gadea, Joaquin; Coy, Pilar; Romar, Raquel; Romero Aguirregomezcorta, Jon; Fisiología
Assisted reproductive technology (ART) is key in cattle breeding as it enables selective mating and improves both reproductive efficiency and genetic improvement. However, there is still limited information on the long-term effects of these methods on the health and development of the offspring. This study aims to investigate the biochemical profile of cattle born through ART, from a previous study (Lopes et al., 2020), over an extended period to understand how these techniques may affect their health and metabolism. Male and female (not pregnant or lactating) cattle born after artificial insemination (AI, N= 7) were compared to those derived from in vitro produced embryos (IVP) using a standard protocol (C-IVP, N= 7), or using oviductal and uterine fluids in the process (RF-IVP, N= 4). Males and females were kept in two diferent open pens, but fed and managed under identical conditions their whole lives. Animals were studied every six months, from 1.5 years old until ≥4 years old, since their first year of life was evaluated by Lopes et al. (2022). Tail vein blood plasma was obtained by centrifugation (1000 G, 10 min) in lithium heparin tubes and stored at 80°C. The biochemical profile included total protein (TP), albumin (ALB), globulin (GLOB), creatinine (CREA), urea (URE), glucose (GLUC), cholesterol (CHOL), triglycerides (TRIG), amylase (AMIL), lipase (LIP), creatinine kinase (CK), alkaline phosphatase (ALP), gamma-glutamyl ranspeptidase (GGT), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and total bilirubin (TB). After serial dilutions, inter- and intraassay precision of the methods were lineal and below 15%. Data normal ly distributed was analysed using ANOVA and Tukey’s test; data not normally distributed was analysed using the Kruskal Wallis and Games Howell’s test (statistical significance p<0.05). The data is shown below as means by group over the years. The AI group had lower CREA and CHOL levels (1.43±0.05 mg/dL and 109.97±5.27 mg/dL, respectively) than C-IVP (1.75±0.07 mg/dL and 126.57±7.53 mg/dL, respectively) and RF-IVP (1.86±0.08 mg/dL and 140.01±7.19 mg/dL, respectively). The GGT levels were lower in RFIVP animals (13.00±1.30 UI/L) than both the C-IVP (17.55±1.10 UI/L) and AI groups (16.05±1.23 UI/L). Moreover, the levels of ALT were significantly lower in C-IVP (28.60±1.72 UI/L) than in the AI (34.35±1.81 UI/ L) and RF-IVP groups (33.40±2.24 UI/L). Depending on the age of the animal at the time of sample collection, significant differences were observed for most parameters. The parameters TP, GLOB, CREA, URE, AMIL, AST an increased in their concentration was observed as the animals aged, whilst ALB, GLUC, ALP decreased with age. However, all values can be considered within physiological ranges despite the significant v ariations. To conclude, the study showed differences in some biochemical parameters amongst the ART groups and with aging. However, despite the diferences all values were within physiological ranges. These findings are key to study the potential implications of this diferences on the animal’s health.
Open Access
Preliminary study of sex hormone levels throughout the estrous cycle in the saliva of 4.5 year old sows born by ART
(2024) Serrano Albal, María; Quintero-Moreno, Armando; Gadea, Joaquín; Romar, Raquel; Romero Aguirregomezcorta, Jon; Anatomía y Anatomía Patológica Comparadas
Estradiol (17-beta-estradiol, E2) and progesterone (P4) play a key role in the regulation of the estrous cycle. Their levels fluctuate throughout the cycle, triggering processes that are essential for the expression of mating behaviour. The rising levels of E2 during the follicular phase triggers the initiation of behavioural and physiological changes associated with heat in sows, whilst P4 is essential for early embryo development and uterine preparation for implantation. The evaluation of sex hormones is typically performed on plasma or serum. However, daily blood collection puts stress on the animals and requires specialized staff, and the extraction may alter the results of some metabolites. Since the small size of these molecules allows them to penetrate the membranes, such as those of the salivary glands, using saliva to measure sex hormones could offer a non-invasive approach to facilitate the collection of sex hormone data. Yet, studies on sex hormones in pig saliva are very scarce, limiting its application in the field. Here, the correlation between the levels of E2 and P4 were evaluated in plasma and saliva, at three separate timepoints, in 22 Large White and Landrace crossbreed sows. These animals were born through different assisted reproductive technologies from a previous study (Paris-Oller et al., 2021), with their ages around 4.5 years old. Their exclusion from production lines allows them to have natural estrous cycles, be kept in an open enclosure, and fed under identical conditions. Furthermore, 12 sows of the same herd [6 born via artificial insemination (AI group) and 6 born after surgical transfer of in vitro-produced embryos (IVP group)] were used to measure the fluctuation of both E2 and P4 in saliva during the estrous cycle. Plasma was obtained through the centrifugation (1000 G, 10 min) of ophthalmic venous sinus blood collected in lithium heparin tubes. Saliva samples were collected using Salivette® tubes containing a polystyrene sponge previously chewed by the sows for 10 secs. These tubes were then centrifuged (1000 G, 5 min) and the samples stored (-80ºC) until hormone analysis. Both plasma and saliva samples were later analysed by quimioluminescence (Immulite). The correlation between the levels of E2 in saliva and plasma, analysed using a Pearson correlation coefficient, showed a statistically significant correlation (p < 0.05) with a moderate negative relationship (-0.311). The average level of plasma E2 was 25.78 pg/mL whilst the saliva E2 was 49.05 pg/mL. However, the correlation between P4 in saliva and plasma was not significant, with an average concentration of 15.23 ng/mL and 0.54 ng/mL in plasma and saliva, respectively. Analyses of the hormone levels in saliva showed no significant differences for both hormone levels between the AI group and IVP group, nor for P4 levels among the days of the estrous cycle. Nevertheless, significant differences were found in the levels of E2 per day of the estrous cycle (p < 0.001). Moreover, in addition to the peak expected towards the end of the follicular phase, another peak was observed around days 12-14 of the cycle. This peak is essential for the maternal recognition of pregnancy in pigs and has been identified in blood but has never been reported in saliva of nonpregnant animals.
Open Access
Growth parameters and growth-related hormone profile in a herd of cattle up to 4 years of age derived from assisted reproductive technologies
(MDPI, 2025-02-21) Heras García, Sonia; Sena Lopes, Jordana; Quintero Moreno, Armando Arturo; Romero Aguirregomezcorta, Jon; Cánovas Bernabé, Sebastián; Romar Andrés, Raquel; Coy Fuster, Pilar; Fisiología; Veterinaria
Assisted reproductive technologies (ART) are routinely used in livestock to generate animals of high genetic value. Despite representing an outstanding accomplishment, recent studies suggest differences in health, fertility, and gestational length of in vitro-produced compared to in vivo-derived animals. Currently, there are no data available on the long-term effects of ART on growth and development. This observational study aimed to understand the relationship between growth and growth-influencing hormones in a herd of cattle derived from artificial insemination (AI) or from in vitro-produced embryos either with BSA (C-IVP) or with reproductive fluids (RF-IVP) as a protein source in culture. Cortisol was associated positively with weight in AI and negatively with body length in males. Thyroxine decreased with age, and it was positively associated with thoracic circumference in RF-IVP. Insulin-like growth factor-1 was greater in RF-IVP than in C-IVP, and it was positively associated with body length and withers height. Growth hormone was greater in females than in males and RF-IVP compared to AI and C-IVP. In conclusion, we present here the first datasets on growth parameters and growth-influencing hormones in cattle from birth to 4 years of age without observing major evidence of differences depending on the embryo origin.
Open Access
Blood plasma biochemical parameters in fully-grown pigs derived from assisted reproductive technologies
(Brazilian College of Animal Reproduction, 2024) Cánovas, Sebastián; Heras, Sonia; Serrano-Albal, María; Ramis, Guillermo; Paris-Oller, Evelyne; Soriano-Úbeda, Cristina; Gadea, Joaquín; Coy, Pilar; Romar, Raquel; Romero Aguirregomezcorta, Jon; Anatomía y Anatomía Patológica Comparadas
Emerging evidence indicates a long-lasting effect of assisted reproductive technologies (ART) on molecular physiology and metabolic function. Previous studies have reported variations in the metabolic profiles in piglets and growing pigs born from artificial insemination (AI) and born after transfer of in vitro produced embryo (IVP) (París-Oller et al., Res Vet Sci,142:43, 2021) but it is unknown whether these differences persist into adulthood. The aim of this work was to study baseline values of plasma biochemical parameters in ART-derived pigs and to gain insight into the evolution of metabolic profile across their life [young age (45 days), adulthood (365 days), and old age (1250 days)]. Pigs born through AI and IVP [n= 16 and 29 (45 days), 13 and 21 (365 days), and 9 and 13 (1250 days), respectively], produced in a previous study (París-Oller et al., J Anim Sci Biotechnol 12:32, 2021), were kept under same housing, managing and feeding conditions. Plasma was obtained through the centrifugation (1200 g, 20 min, 4 ºC, Eppendorf 5810 R) of blood collected in lithium heparin tubes and stored (-80ºC) to determine the biochemical parameters total protein (TP), albumin (ALB), globulin (GLOB), creatinine (CREA), urea (URE), glucose (GLUC), cholesterol (CHOL), triglycerides (TRIG), amylase (AMIL), lipase (LIP), creatinine kinase (CK), alkaline phosphatase (ALP), gamma-glutamyl transpeptidase (GGT), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and total bilirubin (TB) using an automated clinical chemistry analyzer (Olympus AU400, Japan). Inter- and intraassay coefficient of variation were below 15%. Data were analyzed by a mixed ANOVA and Student’s t-test for multiple comparisons. In the absence of homoscedasticity, a Student’s t-test with Welch’s correction was used, and in case of non-normality the Wilcoxon rank-sum test was performed. A P value <0.05 was onsidered significant. The analyses of the metabolites showed higher CREA in old-IVP than old-AI (2.32±0.07 vs. 1.925±0.07 mg/dL); higher GLUC in young-IVP than young-AI (107.77±6.10 vs. 83.77±5.41 mg/dl); lower LIP in young-IVP than young-AI (16.74±1.29 vs. 22.91±2.11 IU/L); and lower AMIL, GGT and AST in IVP than AI at all ages. The other metabolites were similar between AI and IVP animals but increased (PT, ALB, GLOB, URE, TBIL), decreased (CHOL, ALP, ALT) or fluctuated up and down (TRIG) with age. The enzyme CK, related to muscle integrity, was the only parameter that was not affected by the group or age of pigs. Reference values for plasma biochemical values provide valuable information for investigators and will help in valid interpretation for health status and for those who use IVP pigs as a research model. In conclusion, these physiological data are useful for veterinarians and livestock producers and show slight persisting differences in some metabolites in pigs naturally and artificially conceived during life although the clinical relevance of such differences is unnoticeable.