Isolation and characterization of dermal lymphatic and blood endothelial cells reveal stable and functionally specialized cell lineages

J Exp Med. 2001 Sep 17;194(6):797-808. doi: 10.1084/jem.194.6.797.


A plexus of lymphatic vessels guides interstitial fluid, passenger leukocytes, and tumor cells toward regional lymph nodes. Microvascular endothelial cells (ECs) of lymph channels (LECs) are difficult to distinguish from those of blood vessels (BECs) because both express a similar set of markers, such as CD31, CD34, podocalyxin, von Willebrand factor (vWF), etc. Analysis of the specific properties of LECs was hampered so far by lack of tools to isolate LECs. Recently, the 38-kD mucoprotein podoplanin was found to be expressed by microvascular LECs but not BECs in vivo. Here we isolated for the first time podoplanin(+) LECs and podoplanin(-) BECs from dermal cell suspensions by multicolor flow cytometry. Both EC types were propagated and stably expressed VE-cadherin, CD31, and vWF. Molecules selectively displayed by LECs in vivo, i.e., podoplanin, the hyaluronate receptor LYVE-1, and the vascular endothelial cell growth factor (VEGF)-C receptor, fms-like tyrosine kinase 4 (Flt-4)/VEGFR-3, were strongly expressed by expanded LECs, but not BECs. Conversely, BECs but not LECs expressed VEGF-C. LECs as well as BECs formed junctional contacts with similar molecular composition and ultrastructural features. Nevertheless, the two EC types assembled in vitro in vascular tubes in a strictly homotypic fashion. This EC specialization extends to the secretion of biologically relevant chemotactic factors: LECs, but not BECs, constitutively secrete the CC chemokine receptor (CCR)7 ligand secondary lymphoid tissue chemokine (SLC)/CCL21 at their basal side, while both subsets, upon activation, release macrophage inflammatory protein (MIP)-3alpha/CCL20 apically. These results demonstrate that LECs and BECs constitute stable and specialized EC lineages equipped with the potential to navigate leukocytes and, perhaps also, tumor cells into and out of the tissues.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Animals
  • Biomarkers
  • Cell Lineage
  • Cell Separation
  • Cells, Cultured
  • Dermis / blood supply*
  • Endothelium, Vascular / cytology*
  • Endothelium, Vascular / metabolism
  • Extracellular Matrix Proteins / genetics
  • Glycoproteins / biosynthesis
  • Glycoproteins / genetics
  • Humans
  • Hyaluronan Receptors / genetics
  • Intercellular Junctions
  • Lymphatic System / cytology*
  • Lymphatic System / metabolism
  • Membrane Glycoproteins / genetics
  • Mucoproteins / genetics
  • Rabbits
  • Receptor Protein-Tyrosine Kinases / genetics
  • Receptor Protein-Tyrosine Kinases / metabolism
  • Receptor, TIE-2
  • Receptors, Cell Surface / genetics
  • Receptors, Growth Factor / genetics
  • Receptors, Growth Factor / metabolism
  • Receptors, TIE
  • Receptors, Vascular Endothelial Growth Factor
  • Vascular Endothelial Growth Factor Receptor-3
  • Vesicular Transport Proteins


  • Biomarkers
  • Extracellular Matrix Proteins
  • Glycoproteins
  • Hyaluronan Receptors
  • LYVE1 protein, human
  • Membrane Glycoproteins
  • Mucoproteins
  • PDPN protein, human
  • Receptors, Cell Surface
  • Receptors, Growth Factor
  • Vesicular Transport Proteins
  • Receptor Protein-Tyrosine Kinases
  • Receptor, TIE-2
  • Receptors, TIE
  • Receptors, Vascular Endothelial Growth Factor
  • Vascular Endothelial Growth Factor Receptor-3