Isolation, characterization, and comparison of human endometrial and endometriosis cells in vitro.

Abstract

Endometriosis is a common gynecological disorder of unclear pathogenesis. We have established an in vitro model to investigate phenotypic similarities and differences between normal endometrial and endometriosis cells. Highly purified cultures of epithelial and stromal cells were isolated from normal endometrium and endometriosis implants. Morphological features as well as immunocytochemical markers confirm these isolates as epithelial and stromal cells. Potential hormone responsiveness was established by the documentation of estrogen receptor mRNA in epithelial and stromal cells isolated from both tissue types. Expression of this receptor protein was verified in stromal cells by competitive radioligand binding, revealing comparable receptor numbers and dissociation constants. CA-125 is selectively secreted in similar concentrations by epithelial cells isolated from both tissue types. PRL secretion is selectively exhibited by progestin-stimulated stromal cells from both tissue types. Our findings demonstrate that highly purified epithelial and stromal cells cultured from normal endometrial and endometriosis tissues express the same phenotypic and functional markers as their in vivo counterparts. These cultures provide useful models to identify endometriosis-specific cell products that contribute to the pathogenesis of this disorder.

Gap junction communication between uterine stromal cells plays a critical role in pregnancy-associated neovascularization and embryo survival

“…The non-diffusible red DiI marker identifies the microinjected cells. Interestingly, the HESC-T3 cells also failed to undergo morphological decidualization in vitro following treatment with a hormonal cocktail containing E, P and cAMP, whereas the control HESC-TC cells, treated with this cocktail, exhibited distinct epithelioid morphological characteristics (Ryan et al, 1994) that were indicative of their differentiated status (Fig. 7C).…”

“…( C ) HESC-TC and HESC-T3 cells were grown in DMEM/F-12 medium containing 5% charcoal-stripped FBS. To induce in vitro decidualization, they were treated with or without a hormone cocktail containing 1 nM E, 1 mM P and 0.5 mM 8-bromo-cAMP for 7–11 days (Ryan et al, 1994). When the cells were examined morphologically, a distinct transition from fibroblastic to a plump, epitheloid phenotype, characteristic of decidual cells, was observed in control HESC-TC cells.…”

Regulation of vascular endothelial growth factor (VEGF) gene transcription by estrogen receptors α and β

“…Previous studies, by using cytokeratin and vimentin immunostaining, respectively, verified that the epithelial and stromal cell cultures were more than 95% pure. Moreover, the cultures were free of T cells, granulocytes, monocytes, and other leukocytes as detected by CD3, CD11b, and CD45 immunostaining (33).…”

“…All patients had normal ovulatory menstrual cycles and had not received hormonal medication for at least three months at the time of biopsy. Proliferative phase endometrial biopsies were used for the isolation and preparation of endometrial epithelial and stromal cell cultures as described (33). Primary epithelial cells were cultured directly in 24-well dishes.…”

Auf der Spur eines Rätsels: Angiogenese und Endometriose

“…Zwischen ektopischem und eutopischem Endometrium wurden zwar Unterschiede (wie z. B. ein erhöhter Gehalt an Komplement-3-Protein[100], IL-6[104], Tenascin[38] sowie durch IL-1b erhöhte VEGF-Expression[51] in ektopischen Herden) beschrieben, die biochemischen Charakteristiken von Endometriosezellen sind jedoch denen von normalem Endometrium erstaunlich ähnlich[85]. Damit die Gefäûneubildung im Bereich von Endometrioseherden spezifisch angegriffen wird, während das normale Endometrium geschont bleibt, müssen hochspezifische und selektive Medikamente kreiert werden.…”