[Production of soluble intracellular adhesion molecule-1 (sICAM-1) in human endometrial cell culture]

UNLABELLED: Soluble ICAM-1 could play different roles in pathological mechanisms of human endometrium found in such diseases as endometriosis, spontaneous abortion and infertility. The aim of our study was to evaluate in vitro sICAM-1 secretion to medium dependent on menstrual cycle and estradiol (E2) stimulation. MATERIAL AND METHOD: Hysteroscopic biopsies in 19 infertile women. We established 16 cell cultures. Tissue was collected on DMEM, 5% FCS, 5% BSA with Penicillin, Streoptomycin, Gentamycin (2.5 mg/ml). Isolation was performed during 4 hours. Tissue was washed twice, treated with collagenase 1A (1 mg/ml, 37 degrees C 2 hours), centrifuged 10 min by 1200 rpm. Cells were cultured in DMEM/5% FCS/5% BSA with Penicillin/Streptomycin. After 2 days of endometrial and stromal cells coculture D-Valine was used to stop fibroblasts' growth. Number of cells was calculated according to Buerker method. After 2 days estradiol in concentration of 10(-10 M) was added to some cultures. Soluble ICAM-1 concentrations in supernatants were evaluated using standard ELISA kit. T-Student test was used for statistical analysis, p value < 0.05 was considered significant. RESULTS: Median culture time was 14.9 (+/- 3.3) days. Longer cultures were obtained from women in follicular phase. The addition of estradiol prolonged culture by 3.8 days. The level of soluble ICAM-1 in supernatants in follicular phase was 17.6 ng/ml, in luteal 8.8 ng/ml. E2 influenced sICAM-1 production in culture (15.8 vs. 10.6 ng/ml). CONCLUSIONS: Endometrial cells produce sICAM-1 constitutively. Soluble ICAM-1 concentrations are higher in follicular phase. Estradiol increases the excretion of sICAM-1.