title: Transcriptomal comparison of human dermal lymphatic endothelial cells ex vivo and in vitro.
abstract: The in vivo functions of lymphatic endothelial cells depend on their microenvironment, which cannot be fully reproduced in vitro. Because of technical limitations, gene expression in uncultured, "ex vivo" lymphatic endothelial cells has not been characterized at the molecular level. We combined tissue micropreparation and direct cell isolation with DNA chip experiments to identify 159 genes differentiating human lymphatic endothelial cells from blood vascular endothelial cells ex vivo. The same analysis performed with cultured primary cells revealed that only 19 genes characteristic for lymphatic endothelium ex vivo retained this property upon culture, while 27 marker genes were newly induced. In addition, a set of panendothelial genes could be recognized. The propagation of lymphatic endothelial cells in culture stimulated transcription of genes associated with cell turnover, basic metabolism, and the cytoskeleton. On the other hand, there was downregulation of genes encoding extracellular matrix components, signaling via transmembrane tyrosine kinase pathways and the chemokine (C-C) ligand 21. Direct ex vivo analysis of the lymphatic endothelial cell transcriptome is helpful for the understanding of the physiology of the lymphatic vascular system and of the pathogenesis of its diseases.
authors.affiliation: Clinical Institute for Pathology, General Hospital and Medical University of Vienna, Vienna, Austria.
authors.forename: Nikolaus
authors.initials: N
authors.lastname: Wick
authors.forename: Pipsa
authors.initials: P
authors.lastname: Saharinen
authors.forename: Juha
authors.initials: J
authors.lastname: Saharinen
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Gurnhofer
authors.forename: Carl W
authors.initials: CW
authors.lastname: Steiner
authors.forename: Ingrid
authors.initials: I
authors.lastname: Raab
authors.forename: Dejan
authors.initials: D
authors.lastname: Stokic
authors.forename: Pietro
authors.initials: P
authors.lastname: Giovanoli
authors.forename: Sabine
authors.initials: S
authors.lastname: Buchsbaum
authors.forename: Aurea
authors.initials: A
authors.lastname: Burchard
authors.forename: Stefan
authors.initials: S
authors.lastname: Thurner
authors.forename: Kari
authors.initials: K
authors.lastname: Alitalo
authors.forename: Dontscho
authors.initials: D
authors.lastname: Kerjaschki
mesh_terms: D002478:Cells, Cultured; D020405:Dermis / Q000166:cytology / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D004729:Endothelium, Lymphatic / Q000166:cytology / Q000378:metabolism*; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation; D006801:Humans; D007150:Immunohistochemistry; D037181:Lectins, C-Type / Q000235:genetics / Q000378:metabolism; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics; D008957:Models, Genetic; D008962:Models, Theoretical; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic; D000098853:Podoplanin
chemical_list: D037181:Lectins, C-Type; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; C117468:PDPN protein, human; D000098853:Podoplanin