Endometriotic Epithelial Cell Response to Macrophage-Secreted Factors is Dependent on Extracellular Matrix Context
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Endometriosis is a chronic disease in which epithelial and stromal cells that resemble the eutopic endometrium are found in ectopic lesions. In order to examine how microenvironmental factors such as extracellular matrix (ECM) and macrophages influence disease progression, 12Z (an immortalized ectopic epithelial cell line) were cultured on tissue culture plastic or in gels of recombinant basement membrane (rBM) or collagen I. Unlike cells in other conditions, cells in rBM formed multi-cellular structures in a 67 kDa non-integrin laminin receptor (67LR)-dependent manner. To examine the impact of macrophage-secreted factors on cell behavior, 12Z cells on all three substrates were treated with conditioned media from differentiated THP-1 (an immortalized monocytic cell line). Significant proliferation and invasion was observed only with cells cultured in rBM, indicating that ECM cues help dictate cell response to soluble signals. Cells cultured on rBM were then treated with individual cytokines detected in the conditioned media, with increased proliferation observed following exposure to interleukin-8 (CXCL8/IL-8) and both increased proliferation and invasion following treatment with heparin-binding EGF-like growth factor (HB-EGF). This study suggests that rBM gels can be used to induce in vitro lesion formation in order to identify soluble factors that influence proliferation and invasion.
KeywordsEndometriosis 3D culture Heparin-binding EGF-like growth factor (HB-EGF) CXCL8 Basement membrane
We would like to acknowledge Anthony Desotell and Danielle Bourgeois for help with the ErbB ligand ELISAs, Adriana Rodriguez for assistance with the proliferation experiments, Alex LaPerle for providing the ECM adsorption protocols, and the imaging assistance of the Laboratory for Optical and Computational Instrumentation (LOCI) at the University of Wisconsin-Madison. We gratefully acknowledge Dr. Kristyn Masters and Dr. Brenda Ogle for use of the time-lapse microscope. Funding for this work was provided by NSF CBET-0951613 (P.K.K.), American Cancer Society RSG-13-026-01-CSM (P.K.K.), UW-Madison Graduate Research School Grant (P.K.K.), and a NSF GRFP (M.J.C.).
Conflict of interest
Kathryn Pollock, Taylor Jaraczewski, Molly J. Carroll, and Pamela Kreeger declare that they have no conflicts of interest. Dan Lebovic is part of a multi-center endometriosis trial for AbbVie Pharmaceuticals and has received a contribution from UpToDate, Inc.
No human or animal studies were carried out by the authors for this article.
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