Isolation of Human Skin Lymphatic Endothelial Cells and 3D Reconstruction of the Lymphatic Vasculature In Vitro

Methods Mol Biol. 2018;1846:279-290. doi: 10.1007/978-1-4939-8712-2_18.

Abstract

Studies of lymphangiogenesis and lymphatic endothelial biology in vitro require pure cultures of lymphatic endothelial cells and 3D vascular constructs, which closely resemble native human lymphatic vasculature. We describe a method for the isolation of human dermal microvascular lymphatic endothelial cells and generation of a 3D lymphatic capillary network. The lymphatic vascular construct is generated by coculturing primary lymphatic endothelial cells and fibroblasts in their native matrix, without the use of synthetic scaffolds or exogenous factors. The tissue is stable over many weeks and accurately recapitulates features of human dermal lymphatic microvasculature.

Keywords: 3D lymphatic capillary construct; Human lymphatic endothelial cells; Lymphangiogenesis in vitro; Lymphatic endothelial cell isolation; Lymphatic microvascular network in vitro; Self-assembly.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Biomarkers
  • Cell Separation / methods
  • Cells, Cultured
  • Endothelial Cells / metabolism*
  • Endothelium, Lymphatic
  • Extracellular Matrix / metabolism
  • Fibroblasts / metabolism
  • Humans
  • Imaging, Three-Dimensional / methods
  • Immunohistochemistry
  • Lymphatic Vessels / cytology*
  • Lymphatic Vessels / metabolism*
  • Skin / blood supply
  • Skin / metabolism*
  • Workflow

Substances

  • Biomarkers