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Effect of Ca Ionophore On Blastocyst Production Following Intracytoplasmic Sperm Injection in Caprine Oocytes.
Kharche, S D; Pathak, J; Agarwal, S; Kushwah, B; Sikarwar, Aks.
Afiliación
  • Kharche SD; ICAR-CIRG, Makhdoom, Farah, Mathura, UP, India.
  • Pathak J; ICAR-CIRG, Makhdoom, Farah, Mathura, UP, India.
  • Agarwal S; ICAR-CIRG, Makhdoom, Farah, Mathura, UP, India.
  • Kushwah B; ICAR-CIRG, Makhdoom, Farah, Mathura, UP, India.
  • Sikarwar A; ICAR-CIRG, Makhdoom, Farah, Mathura, UP, India.
Reprod Domest Anim ; 51(4): 611-7, 2016 Aug.
Article en En | MEDLINE | ID: mdl-27170442
The aim of the present investigation was to study the effect of calcium ionophore activation on blastocyst production following intracytoplasmic sperm injection (ICSI) in in vitro-matured Caprine oocytes. A total of 470 in vitro-matured oocytes were selected and randomly divided in to three groups. Cumulus oocyte complexes (COCs) recovered by slicing the Caprine ovaries were matured in TCM199 supplemented with 10% foetal bovine serum (FBS) + 10% follicular fluid + FSH (5 µg/ml) + LH (10 µg/ml) + estradiol (1 µg/ml) + EGF (10 ng/ml) + BSA (3 mg/ml) for 27 h in humidified atmosphere at 38.5°C with 5% CO2 in CO2 incubator. After 27 h of culture, selected COCs (n = 470) were separated from cumulus cells by treating with 0.1% hyaluronidase enzyme and passing repeatedly through a fine pipette and randomly divided into three groups. In group 1, (n = 168) matured oocytes were injected with injection micropipette without sperm as control. In group 2, (n = 152) capacitated spermatozoa were injected into cytoplasm of in vitro-matured oocytes through injection micropipette. In group 3, (n = 150) capacitated spermatozoa were injected into cytoplasm of in vitro-matured oocytes through injection micropipette and then activated with 5 µm Ca ionophore for 5 min. The oocytes of all groups were then culture in RVCL media for embryo development. The cleavage rate was observed after 48-72 h of injection. The cleavage rate and blastocyst production in group 1, 2 and 3 were 0.00 and 0.00, 18.42 and 3.57 and 61.33% and 16.30%, respectively. The result indicated that mechanical activation failed to induce cleavage in in vitro-matured Caprine oocytes, whereas chemical activation of intracytoplasmic sperm-injected in vitro-matured Caprine oocytes showed significantly higher cleavage rate and blastocyst production as compare to non-activated oocytes.
Asunto(s)

Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Cabras / Inyecciones de Esperma Intracitoplasmáticas / Ionóforos de Calcio Límite: Animals Idioma: En Revista: Reprod Domest Anim Asunto de la revista: MEDICINA REPRODUTIVA / MEDICINA VETERINARIA Año: 2016 Tipo del documento: Article País de afiliación: India

Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Cabras / Inyecciones de Esperma Intracitoplasmáticas / Ionóforos de Calcio Límite: Animals Idioma: En Revista: Reprod Domest Anim Asunto de la revista: MEDICINA REPRODUTIVA / MEDICINA VETERINARIA Año: 2016 Tipo del documento: Article País de afiliación: India