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1 Clinical Institute for Pathology, General Hospital and Medical University of Vienna, Vienna, Austria
2 Clinical Department of Internal Medicine, Division of Rheumatology, General Hospital and Medical University of Vienna, Vienna, Austria
3 Complex Systems Research Group, Clinical Department of Otorhinolaryngology, General Hospital and Medical University of Vienna, Vienna, Austria
4 Division of Plastic and Reconstructive Surgery, General Hospital and Medical University of Vienna, Vienna, Austria
5 Molecular/Cancer Biology Laboratory, Biomedicum, Helsinki, Finland
6 Department of Molecular Medicine, National Public Health Institute of Finland, and Bioinformatics Unit, Biomedicum, Helsinki, Finland
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.
transcriptome; lymphatic capillary; in vivo
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