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Nerve Growth Factor-Induced Angiogenesis: 1. Endothelial Cell Tube Formation Assay

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Neurotrophic Factors

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1727))

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.

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Acknowledgments

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).

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Authors and Affiliations

  1. Faculty of Medicine, School of Pharmacy, Institute for Drug Research, The Hebrew University of Jerusalem, Jerusalem, Israel

    Philip Lazarovici, Adi Lahiani, Galit Gincberg, Dikla Haham, Arnon Fluksman & Ofra Benny

  2. Department of Bioengineering, College of Engineering, Temple University, Philadelphia, PA, USA

    Cezary Marcinkiewicz & Peter I. Lelkes

Authors
  1. Philip Lazarovici
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  2. Adi Lahiani
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  3. Galit Gincberg
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  4. Dikla Haham
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  5. Arnon Fluksman
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  6. Ofra Benny
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  7. Cezary Marcinkiewicz
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  8. Peter I. Lelkes
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Corresponding author

Correspondence to Philip Lazarovici .

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Editors and Affiliations

  1. Department of Pharmaceutical and Pharmacological Sciences, University of Padova, Padova, Italy

    Stephen D. Skaper

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Lazarovici, P. et al. (2018). Nerve Growth Factor-Induced Angiogenesis: 1. Endothelial Cell Tube Formation Assay. In: Skaper, S. (eds) Neurotrophic Factors. Methods in Molecular Biology, vol 1727. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7571-6_18

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  • DOI: https://doi.org/10.1007/978-1-4939-7571-6_18

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7570-9

  • Online ISBN: 978-1-4939-7571-6

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