2026-06-02T18:28:39Zhttps://docta.ucm.es/rest/oai/request
oai:docta.ucm.es:20.500.14352/1056232024-07-04T23:46:41Zcom_20.500.14352_14col_20.500.14352_15
Docta Complutense author Cañón Beltrán, Karina Esperanza author Cajas Suárez, Yulia Nathaly author González Martínez, María Encina author Fernández-González, Raul author Fierro, Natacha author Lorenzo González, Pedro Luis author Arias Álvarez, María author García García, Rosa María author Gutiérrez Adán, Alfonso author Rizos, Dimitri 2024-07-04T11:35:49Z 2024-07-04T11:35:49Z 2023-03-01 Karina Cañón-Beltrán, Yulia N. Cajas, Encina González, Raúl Fernández-González, Natacha Fierro, Pedro L. Lorenzo, María Arias-Álvarez, Rosa M. García-García, Alfonso Gutiérrez-Adán, Dimitrios Rizos, Acquisition of fertilization competence in guinea pig spermatozoa under different capacitation protocols, Theriogenology, Volume 198, 2023, Pp 231-240, https://doi.org/10.1016/j.theriogenology.2022.12.042. 0093-691X 10.1016/j.theriogenology.2022.12.042 https://hdl.handle.net/20.500.14352/105623 1879-3231 https://doi.org/10.1016/j.theriogenology.2022.12.042 Guinea pig in vitro fertilization (IVF) are poorly developed due to the limited accessibility to oocytes and the lack of an efficient method of sperm capacitation. Thus, we aimed to evaluate different capacitation protocols that we validated through sperm analysis and using heterologous (He) IVF with zona-intact bovine oocytes. Spermatozoa of guinea pigs were collected and processed separately by 4 different protocols: A) Spermatozoa were obtained by flushing the lumen of one cauda epididymis and incubated in a minimal culture medium (MCM); B) One epididymis was placed in a prewarmed of M2 medium and gently minced with fine scissors. Spermatozoa were incubated in a modified human tubal fluid medium (HTF). In both protocols, the spermatozoa were capacitated at 37 °C under an atmosphere of 5% CO2 for 2 h. In the protocols C and D, the spermatozoa were collected by flushing the lumen of the cauda epididymis and selected by commercial density gradient Bovipure® (Nidacon Laboratories AB, Göthenborg, Sweden), according to the manufacturer's instructions. Then for Protocol C) spermatozoa were incubated in MCM medium supplemented with 10 mg/mL heparin (MCM-Hep); while for Protocol D) spermatozoa were incubated in FERT medium supplemented 10 mg/mL heparin (FERT-Hep). Incubation of C and D protocols were performed at 38.5 °C under an atmosphere of 5% CO2 for 2 h. Capacitation protocols C and D showed a higher percentage of viability, total and hyperactive-like motility, and acrosome reaction compared to protocols A and B. For this reason, protocols C and D were used for further He-IVF analysis. Guinea pig sperm and matured zona-intact bovine oocytes were co-incubated at 5% CO2 and 38.5 °C. Sperm-oocyte interaction was assessed at 2.5 h post-insemination (hpi) and pronuclear formation (PrF) were evaluated at 18, 20, 22, 24 and 26 hpi, while the cleavage rate was evaluated at 48 hpi. In protocol D, PrF was significantly higher than in protocol C (P ≤ 0.05) at every time point evaluated. Also, the cleavage rate at 48 hpi was higher (P ≤ 0.05) in He-IVF protocol D (69.8 ± 1.7%) compared to He-IVF protocol C (49.1 ± 1.1%). In conclusion, we determined the most adequate sperm capacitation conditions for guinea pig that allow zona-intact bovine oocyte penetration and lead to hybrid embryo formation, suggesting that these conditions could be optimal to develop IVF in guinea pigs. eng Attribution-NonCommercial-NoDerivatives 4.0 International Acquisition of fertilization competence in guinea pig spermatozoa under different capacitation protocols journal article 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URL https://docta.ucm.es/bitstreams/af817656-869c-4ff3-8e21-5ba0f876fe15/download File MD5 792d421f0c6ac8256b6fd9f34e63a03e 1615925 application/pdf 7ª Cañon_Beltran_Therio_2022.pdf