Cellular composition of primary cultures of human granulosa-lutein cells and the effect of cytokines on cell proliferation

Abstract

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 tumour necrosis factor-alpha (TNF alpha), on the proliferation of individual cell types. After 48 h in culture, most of the cells were granulosa-lutein cells exhibiting positive staining against 3-beta-hydroxysteroid dehydrogenase (3 beta-HSD). About one-third of the total cell population stained positive with monoclonal antibodies against specific antigen sites on lymphohaemopoietic cells, including B-lymphocytes, T-lymphocytes, natural killer cells, monocytes and macrophages. During this 48-h period, IL-2 and TNF alpha significantly (P < 0.05) increased the proportion of leukocyte common antigen (LCA)-positive cells and monocytes. 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 cells. Thus, any observed effects from the addition of cytokines in this system may be due to indirect effects of cytokine-activated leukocytes on granulosa-lutein cells.