[ Objective] The aim of this study was to investigate the effects of bovine follicular microenvironment on the quality of granulosa cells and oocytes as well as Subsequent embryonic development after in vitro fertiliz...[ Objective] The aim of this study was to investigate the effects of bovine follicular microenvironment on the quality of granulosa cells and oocytes as well as Subsequent embryonic development after in vitro fertilization. [ Method] The oocytes and granulosa cells were collected from folli- cles of different diameters. Hoechst33342 staining was used to observe nuclear morphology of oocytes. Annexin-V-FITC/PI staining and flow cytometry were used to detect apoptosis of granulosa cells. After the oosperm were cultured in vitro, fertilization rate and blastocyst rate of oosperm were determined. [Result] The granulosa cells in the large follicles with diameter 〉 10 mm or small follicles with diameter of 0 -3 mm had significantly higher apoptosis rate than those in the follicles with diameter of 3 -7 or 7 -10 mm （P 〈 0.05）. Many oocytes in the follicles with diameter 〉 10 mm or with diameter of 0 -3 mm showed chromatin condensation, nuclear fragmentation and pyknosis. The maturation rate of oocytes with diameter of 3 -7 or 7 -10 mm was significantly higher than that in the large or smell follicles （P 〈 0.05）. The cleavage rate and blastocyst rate of oocytes were not significantly different between the follicles with different diameters. [Condusion~ The follicular microenvironment is a crucial aspect affect- ing in-vitro embryo production. In the follicles with diameter of 3 -10 mm, the quality of granulosa cells as well as the in-vitro maturation rate of oocytes was higher. And the diameter of follicles does not affect oocytes fertilization and early embryonic development of oosperm.展开更多
基金supported by the Science and Technology Fund of Guizhou Province [Guizhou Science and Technology Department,(2008) 2263]
文摘[ Objective] The aim of this study was to investigate the effects of bovine follicular microenvironment on the quality of granulosa cells and oocytes as well as Subsequent embryonic development after in vitro fertilization. [ Method] The oocytes and granulosa cells were collected from folli- cles of different diameters. Hoechst33342 staining was used to observe nuclear morphology of oocytes. Annexin-V-FITC/PI staining and flow cytometry were used to detect apoptosis of granulosa cells. After the oosperm were cultured in vitro, fertilization rate and blastocyst rate of oosperm were determined. [Result] The granulosa cells in the large follicles with diameter 〉 10 mm or small follicles with diameter of 0 -3 mm had significantly higher apoptosis rate than those in the follicles with diameter of 3 -7 or 7 -10 mm （P 〈 0.05）. Many oocytes in the follicles with diameter 〉 10 mm or with diameter of 0 -3 mm showed chromatin condensation, nuclear fragmentation and pyknosis. The maturation rate of oocytes with diameter of 3 -7 or 7 -10 mm was significantly higher than that in the large or smell follicles （P 〈 0.05）. The cleavage rate and blastocyst rate of oocytes were not significantly different between the follicles with different diameters. [Condusion~ The follicular microenvironment is a crucial aspect affect- ing in-vitro embryo production. In the follicles with diameter of 3 -10 mm, the quality of granulosa cells as well as the in-vitro maturation rate of oocytes was higher. And the diameter of follicles does not affect oocytes fertilization and early embryonic development of oosperm.
