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

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Presentation transcript:

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 Nick A. Bersinger, Ph.D., Fioravante Frischknecht, M.D., Robert N. Taylor, M.D., Ph.D., Michael D. Mueller, M.D. Fertility and Sterility Volume 89, Issue 5, Pages 1530-1536 (May 2008) DOI: 10.1016/j.fertnstert.2007.05.075 Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Purified endometrial epithelial glands before distribution into multiwell culture plates. No single (stromal) cells are visible. Bar = 50 μm. Fertility and Sterility 2008 89, 1530-1536DOI: (10.1016/j.fertnstert.2007.05.075) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Immunocytochemistry of endometrial epithelial (left column) and stromal (right column) cells after 4 days in culture. Top row: cytokeratins 5, 6, 8, 17, and 19; middle row: vimentin; bottom row: CD10. Red-brown precipitate reveals positive antigen reaction. Bar = 20 μm. Fertility and Sterility 2008 89, 1530-1536DOI: (10.1016/j.fertnstert.2007.05.075) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Reverse transcriptase polymerase chain reaction of RNA extracted from cultured endometrial epithelial and stromal cells. Amplicon sizes for GAPDH (450 bp) and epithelial neutrophil activating peptide 78 (ENA-78; 162 bp) are indicated by the arrows. Lane 1: 100 bp ladder; lanes 2–5: endometrial epithelial cells (EEC); lanes 6–9: endometrial stromal cells (ESC). Lanes 2 and 6: unstimulated cultures; lanes 3 and 7: interleukin-1β (IL-1β)–stimulated cultures; tracks 4 and 8: tumor necrosis factor α/interferon-γ (TNFα/IFNγ)–stimulated cultures (primers for ENA-78). Lanes 5 and 9: primers for GAPDH (unstimulated cultures). Fertility and Sterility 2008 89, 1530-1536DOI: (10.1016/j.fertnstert.2007.05.075) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 4 Production of ENA-78 (top panel, 11 experiments) and IL-8 (bottom panel, 10 experiments) by cultured endometrial epithelial cells after 8, 24, and 48 hours in culture. The release in the presence of added cytokines is expressed as fold increase over the constitutive release in the absence of cytokines. ∗P<.05; ∗∗P<.001. Abbreviations as in Figure 3. Fertility and Sterility 2008 89, 1530-1536DOI: (10.1016/j.fertnstert.2007.05.075) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 5 Production of ENA-78 (top panel) and IL-8 (bottom panel) by cultured endometrial stromal cells (8 experiments) after 8, 24, and 48 hours in culture in the presence of cytokines (see Fig. 4). ∗P<.05; ∗∗P<.001. Abbreviations as in Figure 3. Fertility and Sterility 2008 89, 1530-1536DOI: (10.1016/j.fertnstert.2007.05.075) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions