In vitro culture of endometrial stromal and gland cells as a model for endometriosis: The effect of peritoneal fluid on proliferation

Objective: To test the hypothesis that the cell-free fraction of PF from women with endometriosis affects the proliferation of endometrial epithelial and stromal cells in vitro. Design: A cell biologic and immunohistochemical study. Setting: University teaching hospital. Patient(s): Premenopausal women undergoing laparoscopy and women with histologically normal endometrium undergoing hysterectomy were selected. Intervention(s): Peritoneal fluid (PF) and serum were collected at laparoscopy. Endometrial epithelial and stromal cells were obtained by enzymic dissociation of tissue, and epithelium was separated from stromal cells by sieving. Epithelial and stromal cell populations were purified by removal of contaminating cells using Thy-1- and CD-45-labeled immunomagnetic beads. Isolated endometrial gland and stromal cells were cultured in the presence of PF or serum from women with and without endometriosis. Main Outcome Measure(s): Cell proliferation was assessed by measurement of incorporation of (3)[H]thymidine after 48 hours in culture. Result(s): Isolated endometrial gland and stromal cells were able to proliferate in vitro. The proliferative effect of PF or sera from women with endometriosis did not differ significantly from normal controls. Conclusion(s): We conclude that PF from women with endometriosis does not have an additional mitogenic effect compared with women without endometriosis. It may be postulated that the endometrium from women with endometriosis responds differently to the effects of PF.