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Hum. Reprod. Advance Access published online on January 4, 2007
Human Reproduction, doi:10.1093/humrep/del491
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© The Author 2007. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Characterization and depletion of leukocytes from cells isolated from the pre-ovulatory ovarian follicle

Péter Fedorcsák1,3, Melinda Ráki2 and Ritsa Storeng1

1 Department of Obstetrics and Gynecology 2 Institute of Immunology, Rikshospitalet-Radiumhospitalet Medical Center, Oslo, Norway

3 To whom correspondence should be addressed at: Department of Obstetrics and Gynecology, Rikshospitalet-Radiumhospitalet Medical Center, 0027 Oslo, Norway. Tel: +47 2307 0219; Fax: +47 2307 2940; E-mail: peter.fedorcsak{at}klinmed.uio.no

BACKGROUND: Cells isolated from the periovulatory ovarian follicle are often used as a model of ovarian steroidogenesis and corpus luteum formation. The follicular fluid-derived cell (FFDC) population is, however, heterogeneous and in addition to granulosa–lutein cells, non-steroidogenic cells are also present. These non-steroidogenic cells, especially the immune cells, may have important biological functions in this model. Here, we describe a method to isolate FFDC, characterize the phenotype of the immune cells and deplete immune cells from FFDC.

METHODS AND RESULTS: Follicular fluid aspirated transvaginally during IVF was clarified by centrifugation and enzymatic dispersion, labelled for leukocyte-specific markers and analysed by flow cytometry. Leukocytes constituted 22% of FFDC and expressed macrophage/dendritic cell, monocyte and lymphocyte markers. Leukocytes were depleted with anti-CD45-conjugated immunobeads, resulting in an FFDC population with <1.9% leukocytes. Leukocyte-containing FFDC secreted more interleukin-8 in culture than leukocyte-depleted FFDC.

CONCLUSION: Leukocyte-depleted FFDC may serve as a useful model to study the interaction of immune cells and luteinizing cells during corpus luteum formation.

Key words: flow cytometry/follicular fluid/granulosa–lutein cells/leukocytes


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