Mutagenesis vol. 19 no. 1 pp. 13-25,
January 2004
© 2004 UK Environmental Mutagen Society/Oxford University Press
Preantral follicle culture as a novel in vitro assay in reproductive toxicology testing in mammalian oocytes
Institute of Gene Technology/Microbiology, Faculty of Biology, University of Bielefeld, D-33501 Bielefeld, Germany and 1Follicle Biology Laboratory, Centre for Reproductive Medicine, University Hospital and Medical School, Free University of Brussels, Laarbeeklaan 101, B-1090 Brussels, Belgium
The most common genetic disorder in humans, trisomy, is caused predominantly by errors in chromosome segregation during oogenesis. Isolated mouse oocytes resuming meiosis and progressing to metaphase II in vitro have recently been used to assess targets, aneugenic potential and sensitivity of oocytes to chemical exposures. In order to extend in vitro maturation tests to earlier stages of oogenesis, an in vitro assay with mouse preantral follicle cultures has been established. It permits the identification of direct and also indirect effects of environmental chemicals on the somatic compartment, the follicle and theca cells, that may lead to disturbances of oocyte growth, maturation and chromosome segregation. Early preantral follicles from prepubertal female mice are cultured in microdroplets for 12 days under strictly controlled conditions. The follicle-enclosed oocytes resume maturation, develop to metaphase II and become in vitro ovulated within 16 h after a physiological ovulatory stimulus with recombinant human gonadotrophins and epidermal growth factor. These oocytes grown and matured in vitro possess normal barrel-shaped spindles with well-aligned chromosomes. Their chromosomes segregate with high fidelity during anaphase I. The model aneugen colchicine induced a meiotic arrest and aneuploidy in these in vitro grown, follicle-enclosed oocytes in a dose-dependent manner, comparable to in vivo tests. Therefore, preantral follicle culture appears to provide an effective and reliable method to assess the influences of environmental mutagens, pharmaceutical agents and potentially endocrine disrupting chemicals on the fidelity of female meiosis.
2To whom correspondence should be addressed. Tel: +49 521 106 6907; Fax: +49 521 106 6015; Email: eiri{at}uni-bielefeld.de
Received on January 9, 2003; revised on August 1 and October 6, 2003; accepted on October 9, 2003
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