The isolation of preantral follicles from the ovaries of bovine was performed under mechanical and enzymatic methods. A significant increase in the total number of follicles retrieved was detected when tissue chopper was used. Micro-dissection could supply good quality, larger sized follicles (400 to ) but with the lowest yield (). The isolated preantral and early antral follicles were cultured for 14 days. Follicles isolated by the mechanical method had a greater growth during a culture period than follicles collected enzymatically. Morphologically normal bovine oocytes from early antral follicles after 14 days culture were 59.6% after culture and after 24 h of maturation culture, 12.9% of in vitro-grown oocytes reached the second metaphase. In conclusion, this study showed that mechanical method can be used effectively to isolate intact preantral follicles from bovine ovaries.