Identification of Angiogenesis Inhibitors Using a Co-culture Cell Model in a High-Content and High-Throughput Screening Platform

SLAS Technol. 2018 Jun;23(3):217-225. doi: 10.1177/2472630317729792. Epub 2017 Sep 18.


Angiogenesis is an important hallmark of cancer, contributing to tumor formation and metastasis. In vitro angiogenesis models for analyzing tube formation serve as useful tools to study these processes. However, current in vitro co-culture models using primary cells have limitations in usefulness and consistency. Therefore, in the present study, an in vitro co-culture assay system was optimized in a 1536-well format for high-throughput screening using human telomerase reverse transcriptase (hTERT)-immortalized mesenchymal stem cells and aortic endothelial cells. The National Center for Advancing Translational Sciences (NCATS) Pharmaceutical Collection (NPC) library containing 2816 drugs was evaluated using the in vitro co-culture assay. From the screen, 35 potent inhibitors (IC50 ≤1 µM) were identified, followed by 15 weaker inhibitors (IC50 1-50 µM). Moreover, many known angiogenesis inhibitors were identified, such as topotecan, docetaxel, and bortezomib. Several potential novel angiogenesis inhibitors were also identified from this study, including thimerosal and podofilox. Among the inhibitors, some compounds were proved to be involved in the hypoxia-inducible factor-1α (HIF-1α) and the nuclear factor-kappa B (NF-κB) pathways. The co-culture model developed by using hTERT-immortalized cell lines described in this report provides a consistent and robust in vitro system for antiangiogenic drug screening.

Keywords: 1536-well plate format; angiogenesis; co-culture cell model; high-content screening.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Angiogenesis Inhibitors / pharmacology*
  • Bortezomib / pharmacology
  • Cell Line, Transformed
  • Coculture Techniques
  • Docetaxel / pharmacology
  • Drug Evaluation, Preclinical / methods*
  • Endothelium, Vascular / drug effects*
  • Endothelium, Vascular / pathology
  • High-Throughput Screening Assays / methods*
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism
  • Mesenchymal Stem Cells / drug effects
  • Mesenchymal Stem Cells / pathology*
  • NF-kappa B / metabolism
  • Neoplasms / drug therapy*
  • Neovascularization, Pathologic / drug therapy*
  • Signal Transduction
  • Telomerase / genetics
  • Topotecan / pharmacology


  • Angiogenesis Inhibitors
  • HIF1A protein, human
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • NF-kappa B
  • Docetaxel
  • Bortezomib
  • Topotecan
  • TERT protein, human
  • Telomerase