title: Development of a combined new mechanical and enzymatic method for the isolation of intact preantral follicles from fetal, calf and adult bovine ovaries.
abstract: 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 mum 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 mum 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.
authors.affiliation: Department of Animal Endocrinology and Reproduction Faculty of Veterinary Medicine University of Liège, B-4000 Liège, Belgium.
authors.forename: J R
authors.initials: JR
authors.lastname: Figueiredo
authors.forename: S C
authors.initials: SC
authors.lastname: Hulshof
authors.forename: R
authors.initials: R
authors.lastname: Van den Hurk
authors.forename: F J
authors.initials: FJ
authors.lastname: Ectors
authors.forename: R S
authors.initials: RS
authors.lastname: Fontes
authors.forename: B
authors.initials: B
authors.lastname: Nusgens
authors.forename: M M
authors.initials: MM
authors.lastname: Bevers
authors.forename: J F
authors.initials: JF
authors.lastname: Beckers