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(18) LEUKEMIA-INHIBITORY FACTOR PROMOTES FOLLICLE SURVIVAL DURING LONG TERM CULTURE OF HUMAN OVARIAN CORTEX. Telfer, Evelyn1, Silfver, Violetta2, Aghajanova, Lusine2, Hovatta, Outi 2, 1 University of Edinburgh, Edinburgh, Scotland2 Karolinska Institute, Stockholm, Sweden ABSTRACT- Leukemia-inhibitory factor (LIF) has previously been shown to promote the primordial to primary follicle transition in rat ovaries (1). The aim of this study was to determine 1) whether LIF could promote primordial initiation during culture of human ovarian cortex and 2) whether LIF protein and LIF-receptor are expressed in the human ovary. Donated ovarian cortical tissue pieces, about 2 x 5 x 5 mm3 in size, were obtained by biopsy from 15 women undergoing gynaecological laparoscopy, after informed consent. The age of the women ranged from 24-42 years. Biopsies were cut into 0.5mm pieces and placed in MEM medium containing BSA, ITS, ascorbic acid, penicillin and streptomycin (control medium) with or without the addition of either 10, 50 or 100ng/ml of human recombinant LIF protein and incubated at 37oC in a humidified incubator with 5% CO2 in air. Medium was changed after 24h and subsequently every 48h. Samples from each treatment group were collected after one and two weeks of culture, fixed in Bouins, and processed for histological examination. Serial sections were stained with Haemotoxylin and Eosin, follicles classified according to stage of development, and oocyte and granulosa cell morphology was scored for signs of degeneration. Control groups were taken for immunocytochemistry for localisation of LIF protein and LIF receptor. The results of the histological analysis showed that LIF treatment had no effect on the proportion of follicles that initiated to grow during either one or two weeks of culture. However, LIF treatment did have a positive effect on the health status of the growing follicle during prolonged culture. After one week of culture there was a significant decrease in oocyte degeneration in the presence of 50 or 100ng/ml of LIF (p>0.05) and a significant reduction in both oocyte and granulosa cell degeneration after two weeks of culture in all concentrations of LIF (p>0.05). Immunocytochemistry showed that LIF protein was localised in granulosa cells and LIF receptor was localised within the oocyte. These results indicate that LIF has a beneficial effect on human follicle survival in vitro and that it may act directly on the oocyte to improve survival. 1) Nilsson, EE, Kezele, P, Skinner, MK., 2002 Mol Cell Endocrinol 188: 65-73 KEY WORDS: culture, Lif-receptor, human ovary, LIF |
