Browsing by Subject "IVF"
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- PublicationOpen AccessAdvanced sperm selection strategies as a treatment for infertile couples: a systematic review(MDPI, 2022-11-10 ) Jordi Ribas-Maynou; Barranco Cascales, Isabel; Sorolla Segura, María; Llavanera, Marc; Delgado Bermúdez, Ariadna; Yeste, Marc; Medicina y Cirugía AnimalAssisted reproductive technology (ART) is an essential tool to overcome infertility, and is a worldwide disease that affects millions of couples at reproductive age. Sperm selection is a crucial step in ART treatment, as it ensures the use of the highest quality sperm for fertilization, thus increasing the chances of a positive outcome. In recent years, advanced sperm selection strategies for ART have been developed with the aim of mimicking the physiological sperm selection that occurs in the female genital tract. This systematic review sought to evaluate whether advanced sperm selection techniques could improve ART outcomes and sperm quality/functionality parameters compared to traditional sperm selection methods (swim-up or density gradients) in infertile couples. According to preferred reporting items for systematic reviews and meta-analyses (PRISMA guidelines), the inclusion and exclusion criteria were defined in a PICOS (population, intervention, comparator, outcome, study) table. A systematic search of the available literature published in MEDLINE-PubMed until December 2021 was subsequently conducted. Although 4237 articles were recorded after an initial search, only 47 studies were finally included. Most reports (30/47; 63.8%) revealed an improvement in ART outcomes after conducting advanced vs. traditional sperm selection methods. Among those that also assessed sperm quality/functionality parameters (12/47), there was a consensus (10/12; 83.3%) about the beneficial effect of advanced sperm selection methods on these variables. In conclusion, the application of advanced sperm selection methods improves ART outcomes. In spite of this, as no differences in the reproductive efficiency between advanced methods has been reported, none can be pointed out as a gold standard to be conducted routinely. Further research addressing whether the efficiency of each method relies on the etiology of infertility is warranted.
- PublicationOpen AccessAldose reductase B1 in pig sperm is related to their function and fertilizing ability(2022-01-31) Mateo Otero, Yentel; Ribas Maynou, Jordi; Delgado Bermúdez, Ariadna; Llavanera, Marc; Recuero, Sandra; Barranco Cascales, Isabel; Yeste, Marc; Medicina y Cirugía AnimalAldose reductase B1 (AKR1B1) has been reported to participate in the modulation of male and female reproductive physiology in several mammalian species. In spite of this, whether or not AKR1B1 could be related to sperm quality, functionality and fertilizing ability is yet to be elucidated. The present study, therefore, aimed to investigate: i) the presence of AKR1B1 in epididymal and ejaculated sperm; ii) the relationship between the AKR1B1 present in sperm and the physiology of the male gamete; iii) the liaison between the relative content of AKR1B1 in sperm and their ability to withstand preservation for 72 h; and iv) the potential link between sperm AKR1B1 and in vitro fertility outcomes. Immunoblotting revealed that AKR1B1 is present in both epididymal and ejaculated sperm with a similar relative content. Moreover, the relative levels of AKR1B1 in sperm (36 kDa band) were found to be negatively related to several kinematic parameters and intracellular calcium levels, and positively to the percentage of sperm with distal cytoplasmic droplets after storage. Finally, AKR1B1 amounts in sperm (36 kDa band) were negatively associated to fertilization rate at two days post-fertilization and embryo development at six days post-fertilization. The results of the present work suggest that AKR1B1 in sperm is probably acquired during maturation rather than at ejaculation and could play a role in that process. Moreover, AKR1B1 seems to be related to the sperm resilience to preservation and to their fertilizing capacity, as lower levels of the 36 kDa band (putative inactive form of this protein) result in better reproductive outcomes.
- PublicationOpen AccessEffect of cumulus cell removal and sperm pre‐incubation with progesterone on in vitro fertilization of equine gametes in the presence of oviductal fluid or cells(2019-05-30) Douet, C; Reigner, F; Goudet, G; Moros Nicolás, Carla; Biología Celular e HistologíaIn spite of many attempts to establish an in vitro fertilization (IVF) technique in the equine, no efficient conventional IVF technique is available. The presence of oviductal fluid or oviductal cells during IVF help to improve embryo production in vitro but is not sufficient to reach high fertilization rates. Thus, our aim was to perform equine IVF either after sperm preincubation with oviductal fluid or in the presence of oviductal cells, and to evaluate the effect of cumulus removal from the oocyte or sperm preincubation with progesterone. In experiment 1 and 2, IVF was performed in the presence of porcine oviduct epithelial cells. The removal of cumulus cells from equine oocytes after in vitro maturation tended to increase the percentage of fertilization when fresh sperm was used (1/33 vs 4/31, p > 0.05) but had no effect when frozen sperm was used (1/32 vs 1/32). Equine sperm preincubation with progesterone did not significantly influence the fertilization rate when fresh or frozen sperm was used (2/14 vs 2/18 for fresh, 1/29 vs 1/25 for frozen). In experiment 3 and 4, IVF was performed after preincubation of sperm with porcine oviductal fluid. The removal of cumulus cells tented to increase the percentage of fertilization when fresh sperm was used (1/24 vs 3/26, p>0.05). Sperm preincubation with progesterone did not significantly influence the fertilization rate when fresh or frozen sperm was used (2/39 vs 2/36 for fresh, 2/37 vs 1/46 for frozen), but two 3-4 cells stage zygotes were obtained with fresh sperm preincubated with progesterone. This is an encouraging result for the setting up of an efficient IVF procedure in equine.
- PublicationOpen AccessEpididymal and ejaculated sperm functionality is regulated differently by periovulatory oviductal fluid in pigs(Wiley, 2020-09-13) Soriano-Úbeda, Cristina; Avilés-López, Karen; García Vázquez, Francisco Alberto; Romero Aguirregomezcorta, Jon; Matas Parra, Carmen; Anatomía y Anatomía Patológica Comparada; Facultad de VeterinariaBackground: The current results of in vitro reproduction techniques in pigs, such as in vitro fertilization (IVF) and embryo development, show high performance with both epididymal and ejaculated spermatozoa. However, the results using ejaculated spermatozoa are even better. Ejaculated spermatozoa are exposed to the secretions of the accessory seminal glands: the seminal plasma (SP). It has been reported that exposure of spermatozoa to reproductive fluids, such as SP or periovulatory oviductal fluid (pOF), modulates sperm functionality both in vivo and in vitro. But whether or not this modulating effect of pOF depends on the origin of the spermatozoa being epididymal or ejaculated, is still unknown. Objectives: To determine and compare the effect of pOF on epididymal and ejaculated sperm functionality. Material and methods: The effects of incubating spermatozoa from the epididymis and ejaculate with pOF in capacitating conditions were investigated by analyzing sperm motility, phosphorylation of protein kinase A substrates and proteins in tyros ine (pPKAs and pTyr, respectively), the interaction of the spermatozoa with the oocyte in IVF and intracytoplasmic sperm injection (ICSI), and, finally, the spermatozoa chromatin condensation status.Results: the pOF modified events related to capacitation in epididymal spermatozoa by decreasing motility, pPKAs and pTyr. In the interaction with the oocyte after sperm capacitation, pOF regulated the epididymal and ejaculated spermatozoa differently. While pOF decreased the number of spermatozoa bound to the zona pellucida (Spz/ZP) and increased oocyte activation after ICSI with epididymal spermatozoa, with the ejaculated spermatozoa, it decreased the mean number penetrating each oocyte (Spz/O). Additionally, pOF significantly increased the nuclear decondensation of the epididymal spermatozoa after the fertilization of the oocyte.Conclusion: The modulation of sperm functionality by pOF is conditioned by the origin of the spermatozoa.
- PublicationRestrictedFertilization outcome could be regulated by binding of oviductal plasminogen to oocytes and by releasing of plasminogen activators during interplay between gametes(2012-02-02) Grullón, LA; Mondéjar Corbalán, Irene; García Vázquez, Francisco Alberto; Romar Andrés, Raquel; Coy Fuster, Pilar; Coy Fuster, Pilar; Fisiología; Facultades de la UMUObjective: To detect plasminogen and plasminogen activators (PA) in oviduct and oocytes and to clarify the role of the plasminogen/plasmin system on mammalian fertilization. Design: Experimental prospective study. Setting: Mammalian reproduction research laboratory. Animal(s): Oviducts and ovaries from porcine and bovine females were collected at slaughterhouse. A total of 52 oviducts and 2,292 oocytes were used. Boar and bull ejaculated spermatozoa were also used. Intervention(s): Plasminogen concentration in oviductal fluid (OF) through the cycle was measured. Immunolocalization of plasminogen and PAs in oocytes was carried out before and after fertilization. Porcine and bovine oocytes were in vitro fertilized, with plasminogen and plasmin added to the culture medium at different concentrations. Main Outcome Measure(s): Plasminogen concentration in OF. Plasminogen and PAs immunolocalization in oocytes. Penetration and monospermy rates, number of spermatozoa in the ooplasma and on the zona pellucida (ZP) after IVF. Result(s): Oviductal fluid contains about 92 mg/mL of plasminogen. The mature oocyte shows immunoreactivity toward plasminogen and toward PAs on its oolemma and ZP. After fertilization, plasminogen and PAs immunolabeling decreases in the oocyte, suggesting its conversion into plasmin. When exogenous plasminogen is added to the IVF medium, sperm entry into the oocyte is hampered, suggesting that the role of plasminogen activation during fertilization is to reduce the number of (or to select) penetrating spermatozoa. Conclusion(s): The plasminogen/plasmin system is activated during gamete interaction and regulates the sperm entry into the oocyte
- PublicationRestrictedHow is plasminogen/plasmin system contributing to regulate sperm entry into the oocyte?(2013-12-30) Grullón LA; Gadea Mateos, Joaquín; Mondéjar Corbalán, Irene; Romar Andrés, Raquel; Matas Parra, Carmen; Coy Fuster, Pilar; Coy Fuster, Pilar; Fisiología; Facultades de la UMUPlasminogen is present in the oviduct, on the zona pellucida (ZP) and on oolemma, and reduces the number of sperm penetrating the oocyte during in vitro fertilization in pig and cow. It is unknown how this reduction occurs. We tested whether plasminogen (1) changed the ZP resistance to enzymatic digestion thus making the passage of the spermatozoa across it difficult; (2) reduced the sperm functionality, assessed by sperm viability, motility, spontaneous acrosome reaction and membrane lipid disorder; or (3) affected the sperm-ZP binding before or after sperm-ZP interaction. The mechanism by which plasminogen/plasmin system contributes to regulate sperm entry into the oocyte is not inducing a ZP hardening or a decrease in sperm functionality but detaching more than 50% of sperm bound to the ZP. It is suggested that the fertilizing spermatozoon activates plasminogen into plasmin at the oocyte surface and that plasmin removes additional spermatozoa attached to the ZP.
- PublicationOpen AccessMimicking the temperature gradient between the sow’s oviduct and uterus improves in vitro embryo culture output(Oxford University Press, 2020-07-09) García Martínez, Soledad; Latorre Reviriego, Rafael Manuel; Sánchez-Hurtado, M. A.; Sánchez-Margallo, F. M.; Bernabò, N.; Romar Andrés, Raquel; López Albors, Octavio Miguel; Coy Fuster, Pilar; Anatomía y Anatomía Patológica ComparadaThis work was designed to determine temperature conditions within the reproductive tract of the female pig and study their impact on ARTs. Temperatures were recorded using a laparo-endoscopic single-site surgery assisted approach and a miniaturized probe. Sows and gilts were used to address natural cycle and ovarian stimulation treatments, respectively. According to in vivo values, IVF was performed at three temperature conditions (37.0°C, 38.5°C and 39.5°C) and presumptive zygotes were cultured in these conditions for 20 h, while further embryo culture (EC) (21–168 h post-insemination) was maintained at 38.5°C. After 20 h, different fertility parameters were assessed. During EC, cleavage and blastocyst stages were evaluated. Sperm membrane fluidity at the experimental temperatures was studied by using differential scanning calorimetry and fluorescence recovery after photobleaching techniques. An increasing temperature gradient of 1.5°C was found between the oviduct and uterus of sows (P < 0.05) and when this gradient was transferred to pig in vitro culture, the number of poly-nuclear zygotes after IVF was reduced and the percentage of blastocysts was increased. Moreover, the temperature transition phase for the boar sperm membrane (37.0°C) coincided with the temperature registered in the sow oviduct, and sperm membranes were more fluid at 37.0°C compared with those of sperm incubated at higher temperatures (38.5°C and 39.5°C). These data suggest that there may be an impact of physiological temperature gradients on human embryo development.