Generation and characterization of a mouse lymphatic endothelial cell line.
Lymphatic vessels, by channeling fluid and leukocytes from the periphery into lymph nodes, play a central role in the development of the immune response. Despite their importance in homeostasis and disease, the difficulties in enriching and culturing lymphatic endothelial cells limit studies of their biology. Here, we report the isolation, stabilization, and characterization of a mouse lymphatic endothelial cell line (MELC) and the generated clones thereof. Cells were isolated from benign lymphangiomas induced by intraperitoneal injections of incomplete Freund's adjuvant. The MELC line expressed molecules typical of lymphatic endothelium, including VEGFR3/Flt-4, podoplanin, Prox-1, and D6, but not LYVE-1. It also expressed CD34, ICAM-1, VCAM, and JAM-A, but not CD31, VE-cadherin, E-selectin, or CX3CL1/fractalkine (both TNFalpha-induced), at variance with vascular endothelial cells tested in parallel. The inflammatory cytokines TNFalpha and IL-4 regulated production of selected adhesion molecules (VCAM), cytokines (IL-6), and chemokines (CCL2/JE). Whole genome transcriptional profiling identified a set of 150 known genes differentially expressed in MELC versus vascular endothelial cells. Thus, the MELC line may represent an invaluable source of lymphatic endothelium.
Department of Immunology and Cell Biology, Mario Negri Institute for Pharmacological Research, Milan, Italy.
Marina
M
Sironi
Annarita
A
Conti
Sergio
S
Bernasconi
Anna M
AM
Fra
Fabio
F
Pasqualini
Manuela
M
Nebuloni
Eleonora
E
Lauri
Maida
M
De Bortoli
Alberto
A
Mantovani
Elisabetta
E
Dejana
Annunciata
A
Vecchi
D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000032:analysis*; D015815:Cell Adhesion Molecules / Q000378:metabolism; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D002999:Clone Cells; D004729:Endothelium, Lymphatic / Q000166:cytology*; D005260:Female; D005620:Freund's Adjuvant; D018398:Homeodomain Proteins / Q000378:metabolism; D007150:Immunohistochemistry; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D008202:Lymphangioma / Q000139:chemically induced / Q000473:pathology; D042601:Lymphatic Vessels / Q000166:cytology*; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D008811:Mice, Inbred DBA; D016513:Mice, SCID; D054447:Receptors, CCR10; D011956:Receptors, Cell Surface / Q000378:metabolism; D019707:Receptors, Chemokine / Q000378:metabolism; D025521:Tumor Suppressor Proteins; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism; D040321:Vascular Endothelial Growth Factor Receptor-3 / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays; D000097628:Chemokine Receptor D6; D000098551:Prospero-Related Homeobox 1 Protein; D000098853:Podoplanin
D018952:Antigens, CD34; D014408:Biomarkers, Tumor; D015815:Cell Adhesion Molecules; D005620:Freund's Adjuvant; D018398:Homeodomain Proteins; D018799:Intercellular Adhesion Molecule-1; D008562:Membrane Glycoproteins; D054447:Receptors, CCR10; D011956:Receptors, Cell Surface; D019707:Receptors, Chemokine; D025521:Tumor Suppressor Proteins; D019010:Vascular Cell Adhesion Molecule-1; D040321:Vascular Endothelial Growth Factor Receptor-3; D000097628:Chemokine Receptor D6; D000098551:Prospero-Related Homeobox 1 Protein; C498686:F11r protein, mouse; C490441:Pdpn protein, mouse; D000098853:Podoplanin