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[Objective] The aim of this study was to investigate the effects of bovine follicles microenvironment on the quality of granulosa cells and oocytes as well as subsequent embryos development after in vitro fertilization.[Method] The oocytes and granulosa cells were collected from follicles 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 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 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 small follicles(P 0.05).The cleavage rate and blastocyst rate were not significantly different between the oocytes in the follicles with different diameters.[Conclusion] The follicles microenvironment is a crucial aspect affecting in-vitro embryos 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.
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