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Isolation and characterization of lymphatic microvascular endothelial cells from human tonsils
Author(s) -
Garrafa Emirena,
Alessandri Giulio,
Benetti Anna,
Turetta Daniela,
Corradi Attilio,
Cantoni Anna Maria,
Cervi Edoardo,
Bonardelli Stefano,
Parati Eugenio,
Giulini Stefano Maria,
Ensoli Barbara,
Caruso Arnaldo
Publication year - 2006
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.20537
Subject(s) - lymphatic endothelium , lymphatic system , agglutinin , tonsil , endothelium , palatine tonsil , pathology , biology , cell sorting , microbiology and biotechnology , flow cytometry , anatomy , chemistry , immunology , lectin , medicine , genetics
Human lymphatic endothelial cells (LECs) have isolated prevalently from human derma and tumors. As specialized lymphatic organs within the oropharynx, palatine tonsils are easily obtained and rich in lymphatic venules. Using a two‐step purification method based on the sorting of endothelial cells with Ulex Europaeus Agglutinin 1 (UEA‐1)‐coated beads, followed by purification with monoclonal antibody D2–40, we successfully purified LECs from human palatine tonsils. The LECs were expanded on flasks coated with collagen type 1 and fibronectin for up to 8–10 passages and then analyzed for phenotypic and functional properties. Cultured cells retained the phenotypic pattern of the lymphatic endothelium of palatine tonsils and expressed functional VEGFR‐3 molecules. In fact, stimulation with VEGFR‐3 ligand, the vascular endothelium grow factor C, induced a marked increase in cell proliferation. Similarly to blood endothelial cells (BECs), LECs were able to form tube‐like structure when seeded in Cultrex basement membrane extract. Comparative studies performed on LECs derived from palatine tonsils and iliac lymphatic vessels (ILVs), obtained with the same procedures, showed substantial discrepancies in the expression of various lymphatic markers. This points to the existence of micro‐ and macrovessel‐derived LECs with different phenotypes, possibly involving different biological activities and functions. Palatine tonsil‐ and ILV‐derived LECs may, therefore, represent new models for investigating function and biochemical properties of these lymphatic endothelia. J. Cell. Physiol. 207: 107–113, 2006. © 2005 Wiley‐Liss, Inc.