Producción in vitro de embriones a partir de complejos cúmulos oocitos tipo II en bovinos Bos indicus

Isabel Catalina Vélez, Alfredo Chica, Rodrigo Urrego, Viviana Torres, Claudia Jimenez-Escobar, Jorge Zambrano-Varon

Resumen


La selección morfológica de los complejos cúmulo-oocito (COC) es crucial para la producción de embriones in vitro. Se ha sugerido que COC que muestran signos de atresia poseen capacidad de generar embriones. El objetivo de este trabajo fue evaluar el efecto de la morfología de los COC provenientes de animales Bos indicus con signos de atresia temprana y sin signos de atresia sobre la producción de embriones in vitro. Se clasificaron COC obtenidos de ovarios de faenado en dos grupos: Tipo I (TI): ooplasma homogéneo con ≥ 4 capas de células del cumulo (CC) compactas y Tipo II (TII): ooplasma granular y ≥ 4 capas de CC ligeramente expandidas. Los COC fueron madurados por 24 horas en medio TCM199 y posteriormente fueron fertilizados in vitro durante 18 h. Los presuntos cigotos se cultivaron in vitro por siete días en medio SOF modificado. La calidad embrionaria se determinó por conteo de blastómeras posterior a tinción con Hoechst 33342. Se usó la prueba t para determinar diferencias estadísticas. La tasa de clivaje para los COC, TI (n=220) y TII (n=161), fue 88 ± 4% y 89 ± 8% respectivamente (p>0,05); la tasa de desarrollo embrionario fue 36 ± 7% y 33 ± 8% (p>0,05) respectivamente. El conteo de blastómeras para ambos grupos fue de (TI:101,TII:104) (n=10), (p>0,05). Los resultados de este trabajo permiten concluir que no hay diferencia en la cantidad y calidad de embriones producidos in vitro utilizando COC tipo I o tipo II, sugiriendo que ambas calidades podrían ser usadas en la producción de embriones in vitro a partir de animales Bos indicus.

In vitro embryo production from cumulus–oocytecomplexes type II in Bos indicus cattle

The morphological selection of cumulus-oocyte complexes (COCs) is animportant step for in vitro embryo production. It has been suggested thatCOC showing signs of atresia have the ability to generate embryos. Theobjective of this work was to evaluate the effect of COC morphology fromBos indicus animals with signs of early atresia versus no signs of atresiaon in vitro embryo production. COC were classified in: Type I (TI): homogeneousooplasm with ≥ 4 layers of compact cumulus cells (CC) and Type II(TII): granular ooplasm and ≥ 4 layers of CC slightly expanded. The COCwere matured in vitro for 24 hours in TCM199 medium and subsequentlyfertilized in vitro for 18 h. The suspected zygotes were cultured in vitro forseven days in modified SOFaa medium. Embryonic quality was determinedby blastomeric count following staining with Hoechst 33342. Student testwas used to determine statistical differences for cleavage, blastocyst rateand blastomeric counts between types of COC. The cleavage rate for TI (n= 220) and TII (n = 161) was 88 ± 4 % and 89 ± 8 % respectively (p> 0.05);embryo development rate was 36 ± 7 % and 33 ± 8 % (p>0.05) respectively.The blastomeric count for both groups was 101 and 104 cells for TI and TIIrespectively (n = 10), (p>0.05). These results demonstrate that there is nodifference in the quantity and quality of embryos produced in vitro usingCOC type I or type II, suggesting that both types could be used for bovine invitro embryo production in Bos indicus cows.

Keywords: atresia, competence, cumulus-oocyte complex.

Produção in vitro de embriões de cumulus oócito complexo detipo II de bovinos Bos indicus

A seleção morfológica dos complexos cumulus-oócito (COC) é crucial para a produçãoin vitro de embriões. Relata-se que COC que apresentam sinais de atresia têm a capacidadede gerar embriões. O objetivo deste estudo foi avaliar o efeito morfológico dosCOC de animais Bos indicus com sinais de atresia precoce ou sim sinais de atresia sobrea produção in vitro de embriões. COC obtidos de ovários de animais abatidos foramclassificados em dois grupos: tipo I (TI): ooplasma homogéneo com ≥ 4 camadas decélulas do cúmulos (CC) compactos e Tipo II (TII): ooplasma granular e ≥ 4 camadas CClevemente expandidas. Os COC foram submetidos à maturação por 24 horas em meioTCM199 e depois fertilizados in vitro por 18 h. Os prováveis zigotos foram cultivadosin vitro por sete dias em meio SOFaa modificado. A qualidade embrionária foi determinadapela contagem dos blastômeros após coloração com Hoechst 33342. O teste tfoi utilizado para determinar diferenças estatísticas significativas. A taxa de clivagempara os COC TI (n = 220) e TII (n = 161) foi de 88 ± 4 % e 89 ± 8 %, respectivamente(p>0,05); a taxa de desenvolvimento embrionário foi de 36 ±7 % e 33 ±8 % (p>0,05),respectivamente. E a contagem de blastômeros para ambos os grupos foi (TI: 101,TII: 104) (n = 10) (p>0,05). Com base nos resultados deste trabalho conclui-se quenão existe diferença na quantidade e qualidade dos embriões produzidos in vitro,utilizando COC tipo I ou tipo II, sugerindo que ambas as qualidades podem ser utilizadasna produção in vitro de embriões em animaes Bos indicus.

Palavras-chave: atresia, complexo cumulo-oocito, a concorrência.

http://dx.doi.org/10.21615/cesmvz.12.2.1


Palabras clave


atresia, competencia, complejo cumulo-oocito.

Citas


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