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


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.


CONCLUSIONS
Leukocyte-depleted FFDC may serve as a useful model to study the interaction of immune cells and luteinizing cells during corpus luteum formation.
Fedorcsák
Péter
P
Department of Obstetrics and Gynecology, Rikshospitalet-Radiumhospitalet Medical Center, 0027 Oslo, Norway. peter.fedorcsak@klinmed.uio.no
Ráki
Melinda
M
Storeng
Ritsa
R
D002499:Centrifugation, Density Gradient; D003338:Corpus Luteum / Q000166:cytology; D017077:Culture Media, Conditioned; D005260:Female; D005434:Flow Cytometry; D015571:Follicular Fluid / Q000166:cytology; D006107:Granulosa Cells / Q000166:cytology; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism; D017493:Leukocyte Common Antigens / Q000096:biosynthesis; D007962:Leukocytes / Q000166:cytology*; D006080:Ovarian Follicle / Q000166:cytology*; D010060:Ovulation; D010641:Phenotype
D017077:Culture Media, Conditioned; D016209:Interleukin-8; D017493:Leukocyte Common Antigens