[en] The isolation of preantral follicles from the ovaries of bovine fetuses, calves and adult cows was performed using a simple, rapid mechanical and enzyme method. The ovaries were cut into small pieces with a tissue chopper. Then, the suspension was filtered successively through 500 and 100 μm nylon mesh filters. This simple mechanical procedure resulted in large numbers of isolated preantral follicles: 2,142 ± 254; 512 ± 92 and 298 ± 54 from the ovaries of bovine fetuses, calves and cows, respectively. In addition, the ovarian fragments between 100 and 500 μm were suspended in 10 ml of M199 Hepes medium plus 5% FCS and divided into 2 equal parts: one portion was used for collagenase treatment (200 U/ml) for 20 minutes, while the other served as a control. Collagenase treatment resulted in 841 ± 161; 216 ± 51 and 52 ± 17 preantral follicles from fetuses, calves and cows, respectively, compared with 312 ± 86; 52 ± 15 and 10 ± 2 in the control group. The use of collagenase with ovarian fragments selected by filtration as a method for increasing the rate of recovery of preantral follicles is described here.
Laboratoire d'Endocrinologie et de Physiologie de la Reproduction animale
I.R.S.I.A. et FRSM et Bourse CNPq-BIOAGRO (UFV) Brésil