Modeling human tumor angiogenesis in a three-dimensional culture system.

Giorgio Seano, Giulia Chiaverina, Paolo Armando Gagliardi, Laura di Blasio, Roberto Sessa, Federico Bussolino, Luca Primo

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

The intrinsic complexity of the process of vessel formation limits the efficacy of cellular assays for elucidation of its molecular and pharmacologic mechanisms. We developed an ex vivo three-dimensional (3D) assay of sprouting angiogenesis with arterial explants from human umbilical cords. In this assay, human arterial rings were embedded in basement membrane extract gel, leading to a network of capillarylike structures upon vascular endothelial growth factor (VEGF) A stimulation. The angiogenic outgrowth consisted of endothelial cells, which actively internalized acetylated-low-density lipoprotein, surrounded by pericytes. Computer-assisted quantification of this vascular network demonstrated considerable sensitivity of this assay to several angiogenic inhibitors, including kinase inhibitors and monoclonal antibodies. We also performed targeted gene knockdown on this model by directly infecting explanted umbilical arteries with lentiviruses carrying short-hairpin RNA. Downregulation of VEGFR2 resulted in a significant reduction of the sprouting capability, demonstrating the relevance of human vascular explants for functional genomics studies. Furthermore, a modification of this assay led to development of a 3D model of tumor-driven angiogenesis, in which angiogenic outgrowth was sustained by spheroids of prostate cancer cells in absence of exogenous growth factors. The human arterial ring assay bridges the gap between in vitro endothelial cell and animal model, and is a powerful system for identification of genes and drugs that regulate human angiogenesis.

Original languageEnglish
JournalBlood
Volume121
Issue number21
DOIs
Publication statusPublished - May 23 2013

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Tumors
Assays
Neoplasms
Endothelial cells
Blood Vessels
Endothelial Cells
Gene Knockdown Techniques
Genes
Pericytes
Lentivirus
Umbilical Arteries
Angiogenesis Inhibitors
Umbilical Cord
Genomics
Basement Membrane
Vascular Endothelial Growth Factor A
Small Interfering RNA
Prostatic Neoplasms
Intercellular Signaling Peptides and Proteins
Phosphotransferases

ASJC Scopus subject areas

  • Hematology
  • Biochemistry
  • Cell Biology
  • Immunology

Cite this

Modeling human tumor angiogenesis in a three-dimensional culture system. / Seano, Giorgio; Chiaverina, Giulia; Gagliardi, Paolo Armando; di Blasio, Laura; Sessa, Roberto; Bussolino, Federico; Primo, Luca.

In: Blood, Vol. 121, No. 21, 23.05.2013.

Research output: Contribution to journalArticle

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