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  • Basal and cytokine-stimulated production of epithelial neutrophil activating peptide-78 (ENA-78) and interleukin-8 (IL-8) by cultured human endometrial epithelial and stromal cells.

Basal and cytokine-stimulated production of epithelial neutrophil activating peptide-78 (ENA-78) and interleukin-8 (IL-8) by cultured human endometrial epithelial and stromal cells.

Fertility and sterility (2007-09-01)
Nick A Bersinger, Fioravante Frischknecht, Robert N Taylor, Michael D Mueller
RESUMEN

To analyze the constitutive production of epithelial neutrophil activating peptide 78 (ENA-78) and interleukin-8 (IL-8) by epithelial cells and the response of these cells to cytokine stimulation. In vitro study using eutopic endometrial tissue. University hospital. Cycling women undergoing laparoscopy for reasons of infertility or unexplained abdominal pain. Isolation of epithelial and stromal cells from endometrium, immunocytochemical characterization and separate culture of these cells in presence of IL-1, tumor necrosis factor alpha (TNF-alpha), and interferon-gamma. Quantitation of IL-8 and ENA-78 released into the medium by ELISA. Polymerase chain reaction was used to demonstrate the presence of ENA-78 in the cell lysate. High purity of the endometrial epithelial cell preparation before culture was demonstrated by the lack of immunocytochemical staining for CD10. Stromal cell preparations were CD10 positive and cytokeratin negative. Stromal cells produced ENA-78 and IL-8 under cytokine stimulation, and epithelial cells were found not only to produce these markers in the absence of cytokine stimulation, but also to increase this output in the presence of IL-1beta or of TNF-alpha plus interferon-gamma. This response may be an important angiogenic step in the early stages in the pathogenesis of endometriosis.

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Sigma-Aldrich
ADN polimerasa Taq from Thermus aquaticus, with 10× PCR reaction buffer without MgCl2