Three-dimensional levitation culture improves in-vitro growth of secondary follicles in bovine model.

RESEARCH QUESTION: Does a three-dimensional culture system based on magnetic levitation with nanoparticles assembly maintain the follicular structure and viability with adequate growth rates leading to oocyte maturation after long-term culture? DESIGN: Randomized-controlled trial of treatments in a bovine model. Secondary follicles (n = 213) isolated from bovine ovaries were cultured in a two-dimensional system (two-dimensional control) or three-dimensional levitation system with different concentrations (three-dimensional 50 µl/ml, 100 µl/ml and 200 µl/ml) of magnetic nanoparticles. Follicular growth (diameter, daily growth and growth patterns), morphology (normal, degenerated and extruded follicles), antrum formation, oocyte viability and chromatin configuration were assessed. RESULTS: Secondary follicles of three-dimensional 200-µl/ml treatment showed higher viability, antrum formation and lower degeneration rates than two-dimensional control. Also, follicles cultured in the three-dimensional 200-µl/ml treatment presented a most homogenous daily growth rate as shown by the lowest variance and standard deviation. Compared with the two-dimensional control, the proportion of non-growing and slow-growing follicles were 3.8-fold lower and 1.6-fold higher, respectively, in the three-dimensional 200-µl/ml treatment. After in-vitro maturation, the three-dimensional 200-µl/ml had a greater proportion of viable oocytes (1.7-fold) and meiotic resumption rates (2.4-fold) than the two-dimensional control treatment. CONCLUSION: The three-dimensional levitation culture system improves the viability of in-vitro development of bovine secondary follicles, antrum formation and lower extrusion and degeneration rates and adequate growth rate leading to relevant oocyte viability and meiotic resumption after in-vitro maturation. This approach does not require a specific medium, and has the potential as an alternative method to in-vitro follicle culture in several species, including humans.

Gonadorelin at the outset and/or end of an ovulation synchronization progesterone estradiol benzoate-based protocol in Nellore females / Gonadorelina no início e/ou no final do protocolo de sincronização da ovulação a base de progesterona e benzoato de estradiol em fêmeas Nelore

This study aimed to evaluate the effects of gonadorelin (GnRH), used both at the outset of a timed artificial insemination (TAI) protocol to synchronize follicular wave recruitment and at the end to synchronize ovulation, on the conception rate (CR) in Nellore cows. The experiment was conducted on two beef cattle farms in the Vale do Araguaia, MT. The body condition score (BCS) evaluation and ultrasound examination to determine ovarian status [follicle diameter < 10 mm, ≥ 10 mm or presence of corpus luteum (CL)] were performed at the beginning (Day 0) of the TAI protocol. Multiparous cows (n = 494) were subjected to the following protocol: Day 0 insertion of intravaginal progesterone device (P4 DIB®, MSD) previously used for 8 or 16 days and IM administration of 2.0 mg of estradiol benzoate (EB, Gonadiol®, MSD); Day 8 P4 device withdrawal, IM injection of 300 IU of equine chorionic gonadotropin (eCG, Folligon®, MSD), 1.0 mg of estradiol cypionate (ECP®, Zoetis) and 0.265 mg of Sodium cloprostenol (PGF2α, Ciosin®, MSD); Day 10 TAI, performed by a single inseminator. Animals were randomly assigned to one of four groups: control (n = 126), GnRH D0 (n = 123), GnRH D10 (n = 123) and GnRH D0 + D10 (n = 122). Animals treated with GnRH received 50 mcg of gonadorelin (GnRH, Cystorelin®, Merial). Pregnancy was diagnosed by ultrasound 39 ± 10 days after TAI. Data were analyzed using the GLIMMIX procedure of SAS program (P < 0.05). The overall CR was 42.71%. Effect of treatment was not detected (P = 0.2482) on CR, which was 36.89% ± 0.19 (47/126) for the control group, 46.08% ± 0.19 (56/123) for GnRH D0, 48.30% ± 0.18 (61/123) for GnRH D10 and 38.48% ± 0.19 (47/122) for GnRH D0 + D10. An effect of farm was detected; the CR was 47.32% ± 0.13 at Farm A and was 38.24% ± 0.13 at Farm B (P = 0.0249). No effects of the interaction between farm and treatment were observed (P = 0.7662), nor of those among ovarian condition and treatment (P = 0.1225) on CR.[...](AU)

Objetivou-se avaliar os efeitos da gonadorelina (GnRH) utilizada para sincronização da emergência da onda de crescimento folicular e para indução da ovulação sincronizada ou em ambos sobre a taxa de concepção (TC) de vacas da raça Nelore. O experimento foi realizado em duas fazendas no Vale do Araguaia, MT. A avaliação do escore de condição corporal (ECC) e a ultrassonografia para determinação da condição dos ovários [folículo com diâmetro < 10 mm, ≥ 10 mm ou presença de corpo lúteo (CL)] foram realizadas no início (Dia 0) do protocolo de inseminação artificial em tempo fixo (IATF). Vacas multíparas (n = 494) foram submetidas ao protocolo: Dia 0 inserção do dispositivo intravaginal de progesterona (P4, DIB®, MSD) utilizado previamente por oito ou 16 dias e aplicação IM de 2,0 mg de Benzoato de Estradiol (BE, Gonadiol®, MSD); Dia 8 remoção do dispositivo, aplicação IM de 300 UI de Gonadotrofina Coriônica Equina (eCG, Folligon®, MSD), de 1,0 mg de Cipionato de Estradiol (ECP®, Zoetis) e de 0,265 mg de Cloprostenol Sódico (PGF2α, Ciosin®, MSD); Dia 10 IATF, realizada por um único inseminador. Os animais foram distribuídos aleatoriamente em quatro grupos: controle (n = 126), GnRH D0 (n = 123), GnRH D10 (n = 123) e GnRH D0 + D10 (n = 122), sendo que os animais tratados com GnRH receberam 50 mcg de gonadorelina (GnRH, Cystorelin®, Merial). O diagnóstico de gestação foi realizado por ultrassonografia 39 ± 10 dias após a IATF. As variáveis foram analisadas pelo procedimento GLIMMIX do programa SAS (P < 0,05). Verificou-se uma TC geral de 42,71%. Não foi detectado efeito dos tratamentos sobre a TC (P = 0,2482), que foi de 36,89% ± 0,19 (47/126) para o grupo controle, 46,08% ± 0,19 (56/123) para GnRH D0, 48,30% ± 0,18 (61/123) para GnRH D10 e de 38,48% ± 0,19 (47/122) para GnRH D0 + D10. Observou-se efeito da variável fazenda, sendo que na Fazenda A a TC foi de 47,32% ± 0,13 e na Fazenda B de 38,24% ± 0,13 (P = 0,0249).[...](AU)

Gonadorelin at the outset and/or end of an ovulation synchronization progesterone estradiol benzoate-based protocol in Nellore females / Gonadorelina no início e/ou no final do protocolo de sincronização da ovulação a base de progesterona e benzoato de estradiol em fêmeas Nelore

This study aimed to evaluate the effects of gonadorelin (GnRH), used both at the outset of a timed artificial insemination (TAI) protocol to synchronize follicular wave recruitment and at the end to synchronize ovulation, on the conception rate (CR) in Nellore cows. The experiment was conducted on two beef cattle farms in the Vale do Araguaia, MT. The body condition score (BCS) evaluation and ultrasound examination to determine ovarian status [follicle diameter < 10 mm, ≥ 10 mm or presence of corpus luteum (CL)] were performed at the beginning (Day 0) of the TAI protocol. Multiparous cows (n = 494) were subjected to the following protocol: Day 0 insertion of intravaginal progesterone device (P4 DIB®, MSD) previously used for 8 or 16 days and IM administration of 2.0 mg of estradiol benzoate (EB, Gonadiol®, MSD); Day 8 P4 device withdrawal, IM injection of 300 IU of equine chorionic gonadotropin (eCG, Folligon®, MSD), 1.0 mg of estradiol cypionate (ECP®, Zoetis) and 0.265 mg of Sodium cloprostenol (PGF2α, Ciosin®, MSD); Day 10 TAI, performed by a single inseminator. Animals were randomly assigned to one of four groups: control (n = 126), GnRH D0 (n = 123), GnRH D10 (n = 123) and GnRH D0 + D10 (n = 122). Animals treated with GnRH received 50 mcg of gonadorelin (GnRH, Cystorelin®, Merial). Pregnancy was diagnosed by ultrasound 39 ± 10 days after TAI. Data were analyzed using the GLIMMIX procedure of SAS program (P < 0.05). The overall CR was 42.71%. Effect of treatment was not detected (P = 0.2482) on CR, which was 36.89% ± 0.19 (47/126) for the control group, 46.08% ± 0.19 (56/123) for GnRH D0, 48.30% ± 0.18 (61/123) for GnRH D10 and 38.48% ± 0.19 (47/122) for GnRH D0 + D10. An effect of farm was detected; the CR was 47.32% ± 0.13 at Farm A and was 38.24% ± 0.13 at Farm B (P = 0.0249). No effects of the interaction between farm and treatment were observed (P = 0.7662), nor of those among ovarian condition and treatment (P = 0.1225) on CR.[...]

Objetivou-se avaliar os efeitos da gonadorelina (GnRH) utilizada para sincronização da emergência da onda de crescimento folicular e para indução da ovulação sincronizada ou em ambos sobre a taxa de concepção (TC) de vacas da raça Nelore. O experimento foi realizado em duas fazendas no Vale do Araguaia, MT. A avaliação do escore de condição corporal (ECC) e a ultrassonografia para determinação da condição dos ovários [folículo com diâmetro < 10 mm, ≥ 10 mm ou presença de corpo lúteo (CL)] foram realizadas no início (Dia 0) do protocolo de inseminação artificial em tempo fixo (IATF). Vacas multíparas (n = 494) foram submetidas ao protocolo: Dia 0 inserção do dispositivo intravaginal de progesterona (P4, DIB®, MSD) utilizado previamente por oito ou 16 dias e aplicação IM de 2,0 mg de Benzoato de Estradiol (BE, Gonadiol®, MSD); Dia 8 remoção do dispositivo, aplicação IM de 300 UI de Gonadotrofina Coriônica Equina (eCG, Folligon®, MSD), de 1,0 mg de Cipionato de Estradiol (ECP®, Zoetis) e de 0,265 mg de Cloprostenol Sódico (PGF2α, Ciosin®, MSD); Dia 10 IATF, realizada por um único inseminador. Os animais foram distribuídos aleatoriamente em quatro grupos: controle (n = 126), GnRH D0 (n = 123), GnRH D10 (n = 123) e GnRH D0 + D10 (n = 122), sendo que os animais tratados com GnRH receberam 50 mcg de gonadorelina (GnRH, Cystorelin®, Merial). O diagnóstico de gestação foi realizado por ultrassonografia 39 ± 10 dias após a IATF. As variáveis foram analisadas pelo procedimento GLIMMIX do programa SAS (P < 0,05). Verificou-se uma TC geral de 42,71%. Não foi detectado efeito dos tratamentos sobre a TC (P = 0,2482), que foi de 36,89% ± 0,19 (47/126) para o grupo controle, 46,08% ± 0,19 (56/123) para GnRH D0, 48,30% ± 0,18 (61/123) para GnRH D10 e de 38,48% ± 0,19 (47/122) para GnRH D0 + D10. Observou-se efeito da variável fazenda, sendo que na Fazenda A a TC foi de 47,32% ± 0,13 e na Fazenda B de 38,24% ± 0,13 (P = 0,0249).[...]