Isolation and characterization of human esophageal microvascular endothelial cells: mechanisms of inflammatory activation

Abstract

Gastroesophageal reflux disease is the most common malady of the esophagus, affecting 7% of the United States population. Histological assessment demonstrates classic inflammatory mechanisms including selective leukocyte recruitment and hemorrhage, suggesting a prominent role for the microvasculature. We isolated and characterized human esophageal microvascular endothelial cells (EC) (HEMEC), examined inflammatory activation in response to cytokines, LPS, and acidic pH exposure, and identified signaling pathways that underlie activation. HEMEC displayed characteristic morphological and phenotypic features including acetylated LDL uptake. TNF-alpha/LPS activation of HEMEC resulted in upregulation of the cell adhesion molecules (CAM) ICAM-1, VCAM-1, E-selectin, and mucosal addressin CAM-1 (MAdCAM-1), increased IL-8 production, and enhanced leukocyte binding. Both acid and TNF-alpha/LPS activation lead to activation of SAPK/JNK in HEMEC that was linked to VCAM-1 expression and U-937 leukocyte adhesion. Expression of constitutive inducible nitric oxide synthase in HEMEC was in marked contrast to intestinal microvascular endothelial cells. In this study, we demonstrate that HEMECs are phenotypically and functionally distinct from lower gut-derived endothelial cells and will facilitate understanding of inflammatory mechanisms in esophageal inflammation.

Endothelial-mesenchymal transition in normal human esophageal endothelial cells cocultured with esophageal adenocarcinoma cells: role of IL-1β and TGF-β2

“…The use of human tissue, cells, and all experiments were approved by the Institutional Review Board of the Medical College of Wisconsin. Primary HEMEC were isolated as described previously (22). Endothelial cultures were recognized by modified lipoprotein uptake (Dil-ac-LDL; Life Technologies, Grand Island, NY) and expression of Factor VIII associated antigen.…”

“…Endothelial cultures were recognized by modified lipoprotein uptake (Dil-ac-LDL; Life Technologies, Grand Island, NY) and expression of Factor VIII associated antigen. Primary human intestinal microvascular endothelial cells (HIMEC) were isolated from surgical tissue as described previously (22).…”

“…Paraffin-embedded EAC specimens (5-m sections) were obtained from the Department of Pathology, Medical College of Wisconsin and stained with hematoxylin-eosin for histological diagnosis. The sections were subjected to immunofluorescence staining using the above antibodies, as described previously (22).…”

“…SDS-PAGE and imunoblot analysis were performed as described previously (22), using the antibodies mentioned above.…”

Antibody Library-based Tumor Endothelial Cells Surface Proteomic Functional Screen Reveals Migration-stimulating Factor as an Anti-angiogenic Target

“…Isolation of Endothelial Cells from Human Tissues-Human esophageal cancer endothelial cells (HECECs) were isolated from esophageal squamous cell carcinoma tissues (12,13). Briefly, to isolate pure endothelial cell population from human esophageal squamous cell carcinoma, cancerous tissues were obtained Ͻ30 min after surgical removal.…”