CELLULAR COMPOSITION OF PRIMARY CULTURES OF HUMAN GRANULOSA-LUTEIN CELLS AND THE EFFECT OF CYTOKINES ON CELL-PROLIFERATION

The cellular composition of cells collected from the follicular fluid obtained during the IVF procedure and cultured in vitro was examined, as well as the effects of two cytokines, interleukin-2 (IL-2) and tumo ur necrosis factor-alpha (TNF alpha), on the proliferation of individu al cell types. After 48 h in culture, most of the cells were granulosa -lutein cells exhibiting positive staining against 3-beta-hydroxystero id dehydrogenase (3 beta-HSD). About one-third of the total cell popul ation stained positive with monoclonal antibodies against specific ant igen sites on lymphohaemopoeitic cells, including B-lymphocytes, T-lym phocytes, natural killer cells, monocytes and macrophages. During this 48-h period, IL-2 and TNF alpha significantly (P < 0.05) increased th e proportion of leukocyte common antigen (LCA)-positive cells and mono cytes. At the end of 144 h in culture, although LCA-positive cells and monocytes were still present, there were fewer. The main targets for the proliferative effects of IL-2 and TNF alpha in this culture system during the first 48 h are leukocytes rather than steroid-producing ce lls. Thus, any observed effects from the addition of cytokines in this system may be due to indirect effects of cytokine-activated leukocyte s on granulosa-lutein cells.