Intra-laboratory validated human cell-based in vitro vasculogenesis/angiogenesis test with serum-free medium

Reprod Toxicol. 2017 Jun;70:116-125. doi: 10.1016/j.reprotox.2016.11.015. Epub 2016 Nov 30.

Abstract

Vasculogenesis and angiogenesis are the processes by which new blood vessels are formed. We have developed a serum-free human adipose stromal cell and umbilical cord vein endothelial cell based vasculogenesis/angiogenesis test. In this study, the test was validated in our GLP laboratory following the OECD Guidance Document 34 [1] using erlotinib, acetylic salicylic acid, levamisole, 2-methoxyestradiol, anti-VEGF, methimazole, and D-mannitol to show its reproducibility, repeatability, and predictivity for humans. The results were obtained from immunostained tubule structures and cytotoxicity assessment. The performance of the test was evaluated using 26 suspected teratogens and non-teratogens. The positive predictive value was 71.4% and the negative predictive value was 50.0%, indicating that inhibition of vasculogenesis is a significant mechanism behind teratogenesis. In conclusion, this test has great potential to be a screening test for prioritization purposes of chemicals and to be a test in a battery to predict developmental hazards in a regulatory context.

Keywords: Angiogenesis; Chemical testing; Human adipose stromal cells; Human umbilical cord vein endothelial cells; In vitro; Teratogens; Vasculogenesis.

Publication types

  • Validation Study

MeSH terms

  • 2-Methoxyestradiol
  • Adipose Tissue / cytology
  • Aspirin / pharmacology
  • Cells, Cultured
  • Culture Media
  • Drug-Related Side Effects and Adverse Reactions
  • Erlotinib Hydrochloride / pharmacology
  • Estradiol / analogs & derivatives
  • Estradiol / pharmacology
  • Human Umbilical Vein Endothelial Cells / drug effects*
  • Human Umbilical Vein Endothelial Cells / physiology
  • Humans
  • Laboratories
  • Levamisole / pharmacology
  • Mannitol / pharmacology
  • Methimazole / pharmacology
  • Neovascularization, Physiologic / drug effects*
  • Reproducibility of Results
  • Serum
  • Stromal Cells / drug effects*
  • Stromal Cells / physiology
  • Teratogens / toxicity*
  • Toxicity Tests / methods*
  • Umbilical Cord / cytology
  • Vascular Endothelial Growth Factor A / antagonists & inhibitors

Substances

  • Culture Media
  • Teratogens
  • Vascular Endothelial Growth Factor A
  • Levamisole
  • Mannitol
  • Estradiol
  • Methimazole
  • 2-Methoxyestradiol
  • Erlotinib Hydrochloride
  • Aspirin