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

Overton, Caroline; Fernández-Shaw, S.; Hicks, B.R.; Barlow, David H.; Starkey, Phyllis M.

doi:10.1016/s0015-0282(97)81855-9

Cited by 21 publications

(11 citation statements)

References 15 publications

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“…Although stimulatory effects of individual substances are obvious, the concept that peritoneal fluid could explain the development of endometriotic disease has never been proven. Indeed, the overall effect in vitro of peritoneal fluid upon the proliferation of purified endometrial stromal and epithelial cells was stimulatory, but no differences were found between women with and without endometriosis [121].…”

Section: Conclusion and Discussionmentioning

confidence: 93%

Pathogenesis of Endometriosis: The Role of Peritoneal Fluid

Koninckx

Kennedy

1999

Gynecol Obstet Invest

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Peritoneal fluid is a specific microenvironment. It originates mainly as an ovarian exudation product due to increased vascular permeability, and contains large amounts of macrophages and their secretion products, which include growth factors, cytokines and angiogenic factors. Similarly, within the ovary, there is also a specific microenvironment, best characterised by the follicular milieu with steroid hormone concentrations that are 1,000 times higher than in plasma. Since endometrial cells in peritoneal fluid and superficially implanted cells will be influenced by peritoneal fluid concentrations, any differences found between minimal endometriosis and eutopic endometrium could be the consequence of their different local environments, rather than inherent cellular differences. Superficial endometrial implants may be regulated by factors in peritoneal fluid, while deep endometriosis and cystic ovarian endometriosis may be under the influence of factors in blood and within the ovary, in which case minimal endometriosis may be physiologic only, while deep endometriosis and cystic ovarian endometriosis, both associated with pelvic pain and infertility, may be a pathological state. In conclusion, the local endocrine environment, that is, in blood, peritoneal fluid, and within the ovary, should be taken into account to explain the differences between superficial, deep and cystic ovarian endometriosis, together with inherent differences in the endometrial cells of women with and without endometriosis.

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Section: Conclusion and Discussionmentioning

confidence: 93%

Pathogenesis of Endometriosis: The Role of Peritoneal Fluid

Koninckx

Kennedy

1999

Gynecol Obstet Invest

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“…Purity of the cell cultures was inferred from the mutually exclusive differential expression of lineage-/tissue-specific genes in the eEC and eSF populations. In addition, we used the well-established conventional markers of eEC and eSF culture purity, KRT18 and vimentin (21, 36, 37), respectively, in both gene expression and protein studies. Independent confirmation of cell-specific expression of KRT18 and vimentin in eEC and eSF, respectively, is provided by our differential gene expression data of highly pure endometrial eEC versus eSF cell populations isolated by FACS using independent cell surface selection markers (Supplemental Table 2).…”

Section: Discussionmentioning

confidence: 99%

“…Gene expression of eSF markers, for example, vimentin and secreted cytokines, was validated at the protein level through immunofluorescence and secreted protein measurements. Although vimentin has been shown to be expressed in a number of cell types, within normal endometrium its stromal localization is well established, and its use as a marker for the identification of eSF in cell cultures is reported in multiple models (21, 36, 37). The expression of PDGFRB is well documented in endometrial stromal fibroblasts, as well as in mesenchymal stem cells, and is indeed used as a selection marker for FACS isolation of these endometrial cell types (28, 64).…”

Section: Discussionmentioning

confidence: 99%

Coculturing human endometrial epithelial cells and stromal fibroblasts alters cell-specific gene expression and cytokine production

Chen

Erikson

Piltonen

et al. 2013

Fertility and Sterility

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Objective To determine the effects of coculturing endometrial epithelial cells (eEC) with paired endometrial stromal fibroblasts (eSF) on cell-specific gene expression and cytokine secretion patterns. Design In vitro study. Setting University research laboratory. Patient(s) Endometrial biopsies were obtained from premenopausal women. Intervention(s) Polarized eEC and subject-paired eSF were cultured for 12.5 hours alone (monoculture) or combined in a two-chamber coculture system without cell-cell contact. Cells and conditioned media were analyzed for global gene expression and cytokine secretion, respectively. Purified, endometrial tissue-derived eEC and eSF isolated by fluorescent activated cell sorting (FACS) were used as noncultured controls. Main Outcome Measure(s) Cell-specific global gene expression profiling and analysis of secreted cytokines in eEC/eSF cocultures and respective monocultures. Result(s) Transepithelial resistance, diffusible tracer exclusion, expression of tight junction proteins, and apical/basolateral vectorial secretion confirmed eEC structural and functional polarization. Distinct transcriptomes of eEC and eSF were consistent with their respective lineages and their endometrial origin. Coculture of eEC with eSF resulted in altered cell-specific gene expression and cytokine secretion. Conclusion(s) This coculture model provides evidence that interactions between endometrial functionally polarized epithelium and stromal fibroblasts affect cell-specific gene expression and cytokine secretion underscoring their relevance when modeling endometrium in vitro.

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“…Several other studies have focused on the effect of PF on endometrial cell viability and proliferation [8][9][10][11]. However, in these studies the proliferative effect of PF from women with and without endometriosis was tested on stromal cells of cyclic and not menstrual endometrium.…”

Section: Discussionmentioning

confidence: 99%

Menstruum Induces Changes in Mesothelial Cell Morphology

Koks

Weusten

Groothuis

et al. 2000

Gynecol Obstet Invest

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In previous studies, we have shown that menstrual endometrium preferentially adheres to the subepithelial lining of the peritoneum. It remains to be elucidated, however, whether this damage is preexisting or inflicted by the menstrual tissue itself. We hypothesized that the menstrual tissue itself damages the peritoneum. To investigate this, the viability of menstrual endometrial tissue in peritoneal fluid (PF) was evaluated and the morphologic changes in the mesothelial cells were studied by in vitro cocultures of menstruum with mesothelial cell monolayers. Menstruum was collected with a menstrual cup. Endometrial tissue was isolated from the menstruum, resuspended in culture medium or in the cell-free fraction of PF and cultured for 24, 48 or 72 h. A 3(4,5-dimethylthia-zolyl-2)-2,5-diphenyl tetrazolium bromide (MTT) assay was performed to obtain a relative measure of viable adhered endometrial cells. Mesothelial cells isolated from human omental tissue were cultured on Matrigel or uncoated plastic. At confluence, overnight cocultures were performed and scanning electron microscopy was used to evaluate the morphologic changes. The viability of endometrial fragments was 84% (n = 36, p < 0.05), 82% (n = 27, not significant) and 104% (n = 14, not significant) when cultured in the cell-free fraction of PF for 24, 48 and 72 h, respectively, when compared to medium with 10% fetal calf serum. Menstrual endometrial fragments or menstrual serum added to and cocultured with mesothelial cells induced severe morphologic alterations of the latter, including retraction, shrinking and gap formation. Similar morphologic changes were observed when mesothelial cells were cocultured with menstrual endometrial fragments in PF or in culture inserts. Incubation with conditioned medium from cultured menstrual endometrium induced similar but less pronounced changes in morphology. In conclusion, menstrual endometrial fragments remain viable in PF in vitro for at least 72 h. Antegradely shed menstruum induces changes in mesothelial cell morphology, including retraction and shrinking with exposure of the underlying surface. These findings suggest that menstruum is harmful to the peritoneal lining. Therefore, by local destruction of the mesothelial layer, menstrual endometrium is able to create sites for adhesion.

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

Cited by 21 publications

References 15 publications

Pathogenesis of Endometriosis: The Role of Peritoneal Fluid

Pathogenesis of Endometriosis: The Role of Peritoneal Fluid

Coculturing human endometrial epithelial cells and stromal fibroblasts alters cell-specific gene expression and cytokine production

Menstruum Induces Changes in Mesothelial Cell Morphology

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