Endometriotic Epithelial Cell Response to Macrophage-Secreted Factors is Dependent on Extracellular Matrix Context

Abstract

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 and macrophages influence disease progression, 12Z (an immortalized ectopic epithelial cell line) were cultured on tissue culture plastic (TCP) 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 extracellular matrix 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.

Impaired Localization of Claudin-11 in Endometriotic Epithelial Cells Compared to Endometrial Cells

“…22 The epithelial cells 12Z and 49Z (provided by Prof. Starzinski-Powitz, Frankfurt, Germany) from active peritoneal endometriotic lesions 23 show characteristics of the active phase of endometriosis and thus are suitable for studying cellular behavior of endometriosis 22 and especially for invasion. 24 The Ishikawa cells originate from an endometrial adenocarcinoma and were purchased from ATCC (Manassas, USA). While this cell line certainly does not represent healthy or endometriotic tissue, it retains characteristics of endometrial glandular epithelium and thus is used in several studies on endometriosis as a representative cell line for the human endometrium.…”

Collagen I triggers directional migration, invasion and matrix remodeling of stroma cells in a 3D spheroid model of endometriosis

“…While the 12Z cell line was created from lesion-derived cells 29 based on their ability to penetrate through Matrigel coated invasion chambers, the 12Z single cells in our study only assembled into cellular aggregates with processes and 12Z spheroids developed invasive edges. Previously, Pollock and colleagues also observed only low levels of basal invasion in 12Z cells on Matrigel hydrogels 45 . We speculate that the limited invasive capacity of 12Z cell observed in this study could be due to the chemotactic gradient that is a key part of the invasion chamber setup.…”

Engineering Approaches to Study Cellular Decision Making

“…A similar dependence of soluble cue signaling on ECM identity has been demonstrated in 3D matrices. For example, endometriotic epithelial cells were responsive to macrophage-conditioned media when cultured on basement membrane, but not on collagen I or TCPS (115). These findings demonstrate the inability of traditional culture platforms (e.g., TCPS) to capture important cellular behaviors and responses.…”