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

  • Anita Rogic
  • Francois Auger
  • Mihaela SkobeEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1846)

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.

Key words

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

Notes

Acknowledgements

This work was supported by the DOD grant BC044819, the Susan G. Komen grant IBC17512321, by the DRPPP Pilot Project Program, Dept. Oncological Sciences, Mount Sinai (M.S.), and the CIHR grant SAC92850 and the Fondation d’entreprise SILAB-Jean Paufique (F.A.).

Supplementary material

Video 1

(MOV 15607 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Oncological Sciences and Tisch Cancer InstituteIcahn School of Medicine at Mount SinaiNew YorkUSA
  2. 2.Centre LOEX de l’Université Laval, Regenerative Medicine Section of the FRQS Research Center of the CHU de QuébecQuebecCanada

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