Effect of fetal age and method of recovery on isolation of preantral follicles from bovine ovaries

The objective of this study was to compare enzymatic and mechanical methods, at distinct fetal ages, on isolation of different developmental stages of preantral follicles from bovine ovaries. Fetal ovaries were obtained from pregnant cattle at 150 to 270 d of gestation, and 135,521 preantral follicl...

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Bibliographic Details
Published in:Theriogenology 1999-09, Vol.52 (4), p.563-571
Main Authors: Carámbula, S.F., Gonçalves, P.B.D., Costa, L.F.S., Figueiredo, J.R., Wheeler, M.B., Neves, J.P., Mondadori, R.G.
Format: Article
Language:English
Subjects:
Animals
bovine
Cattle
Cell Separation - methods
Embryonic and Fetal Development
Female
Fetus
folliculogenesis
Gestational Age
Ovarian Follicle - cytology
Ovarian Follicle - embryology
Ovary - cytology
Ovary - embryology
preantral follicles
Pregnancy
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Summary:The objective of this study was to compare enzymatic and mechanical methods, at distinct fetal ages, on isolation of different developmental stages of preantral follicles from bovine ovaries. Fetal ovaries were obtained from pregnant cattle at 150 to 270 d of gestation, and 135,521 preantral follicles at different stages of development were studied. The dissociation of ovaries with a mechanical procedure resulted in an average of 938.16 preantral follicles. In contrast, 3,715.56 follicles were obtained when enzymatic digestion was used (P = 0.0001). Histological evaluation confirmed follicular stages and demonstrated that both mechanical and mechanical-enzymatic procedure did not affect the cellular integrity of the follicles. Granulosa cell-oocyte complexes surrounded by a basal membrane, were considered preantral follicles in this study. The ratio of different stages of isolated preantral follicles was significantly (P = 0.0001) correlated to fetal age. The earliest fetal age at which tertiary follicles were identified was at 210 d of gestation. The results confirm previous observation that follicular development and atresia are initiated during fetal development. These data provide information on methodologies to isolate intact bovine preantral follicles for investigating the control and regulation of follicular development and the growth of preantral follicles in vitro.
ISSN:0093-691X
1879-3231
DOI:10.1016/S0093-691X(99)00152-1