Morphological changes in mesothelial cells induced by shed menstrual endometrium in vitro are not primarily due to apoptosis or necrosis

A Y Demir Weusten, Patrick G. Groothuis, P G Groothuis, G A Dunselman, A F de Goeij, J W Arends, Jan‐Willem Arends, J.L.H. Evers, J L Evers
Human Reproduction · 2000 · vol. 15(7) , pp. 1462–1468 · doi:10.1093/humrep/15.7.1462 · PMID:10875851 · W2168543980
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Menstrual effluent conditioned media induced significant morphological remodeling in mesothelial cells, without substantial apoptosis or necrosis, potentially facilitating endometriosis development.

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Abstract

In a previous study on the pathogenesis of endometriosis, we observed that constituents of menstrual effluent induce morphological alterations in human mesothelial cells. In this study, we investigated whether these alterations were associated with apoptosis or necrosis or were the result of cellular remodelling. After overnight incubation of confluent monolayers of human omental mesothelial cells (HOMEC) with conditioned media prepared from menstrual effluent shed anterogradely, severe alterations in morphology were observed. Typical polygonal mesothelial cell cultures at confluency acquired elongated spindle morphology, resulting in gaps between the cells. In contrast, mesothelial cells from the control groups receiving culture medium only, retained a normal morphology. Immunofluorescence staining revealed that cytokeratin, vimentin and actin filaments were still present, homogeneously distributed in the cell cytoplasm following changes in morphology. To evaluate whether the morphological alterations were associated with apoptosis and/or necrosis, the cells were stained with the M30 CytoDeath antibody or annexin V with propidium iodide and analysed using flow cytometry. The results showed that only a small percentage (1-7%) of the affected HOMEC were undergoing apoptosis or necrosis. We conclude that the profoundly altered morphology of HOMEC is a result of cellular remodelling and that the role of apoptosis and necrosis is negligible. Soluble paracrine factors released by cells isolated from menstrual effluent shed anterogradely may induce a reorganization of the cytoskeleton. As a result, the underlying basement membrane will be exposed and the mesothelium may no longer prevent implantation of endometrium shed retrogradely into the peritoneum, thus facilitating the development of endometriosis.

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Condition tags

endometriosis

MeSH descriptors

Endometrium Menstruation Omentum Annexin A5 Annexin A5 Apoptosis Cell Line Cells, Cultured Culture Media, Conditioned Culture Media, Conditioned DNA DNA Endometrium Epithelial Cells Epithelial Cells Epithelial Cells Etoposide Etoposide Female Flow Cytometry

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