Nerve growth factor-induced angiogenesis: 1. Endothelial cell tube formation assay

Philip Lazarovici*, Adi Lahiani, Galit Gincberg, Dikla Haham, Arnon Fluksman, Ofra Benny, Cezary Marcinkiewicz, Peter I. Lelkes

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

11 Scopus citations

Abstract

Nerve growth factor (NGF) is a neurotrophin promoting survival, proliferation, differentiation, and neuroprotection in the embryonal and adult nervous system. NGF also induces angiogenic effects in the cardiovascular system, which may be beneficial in engineering new blood vessels and for developing novel anti-angiogenesis therapies for cancer. Angiogenesis is a cellular process characterized by a number of events, including endothelial cell migration, invasion, and assembly into capillaries. In vitro endothelial tube formation assays are performed using primary human umbilical vein endothelial cells, human aortic endothelial cells, and other human or rodent primary endothelial cells isolated from the vasculature of both tumors and normal tissues. Immortalized endothelial cell lines are also used for these assays. When seeded onto Matrigel, these cells reorganize to create tubelike structure, which may be used as models for studying some aspects of in vitro angiogenesis. Image acquisition by light and fluorescence microscopy and/or quantification of fluorescently labeled cells can be carried out manually or digitally, using commercial software and automated image processing. Here we detail materials, procedure, assay conditions, and cell labeling for quantification of endothelial cell tube formation. This model can be applied to study cellular and molecular mechanisms by which NGF or other neurotrophins promote angiogenesis. This model may also be useful for the development of potential angiogenic and/or anti-angiogenic drugs targeting NGF receptors.

Original languageAmerican English
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages239-250
Number of pages12
DOIs
StatePublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1727
ISSN (Print)1064-3745

Bibliographical note

Funding Information:
Philip Lazarovici holds the Jacob Gitlin Chair in Physiology and is affiliated and supported by the David R Bloom Center for Pharmacy and the Adolph and Klara Brettler Medical Research Center at the Hebrew University of Jerusalem, Israel. Peter I. Lelkes is the Laura H. Carnell Professor of Bioengineering and Chair of the Department of Bioengineering, Temple University. Research reported in this publication was supported in part by a grant from the Temple University Moulder Center for Drug Discovery (CM and PIL) and a Research Bridge Funding Award (PIL) from the Temple University Office of the Vice President for Research Administration (OVPR).

Publisher Copyright:
© Springer Science+Business Media, LLC 2018.

Keywords

  • Analysis
  • Angiogenesis
  • Endothelial cellw tube formation assay
  • Matrigel
  • Nerve growth factor

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