An Endothelial Cell/Mesenchymal Stem Cell Coculture Cord Formation Assay to Model Vascular Biology In Vitro

Methods Mol Biol. 2018:1683:371-382. doi: 10.1007/978-1-4939-7357-6_21.

Abstract

Blood vessels are crucial components for normal tissue development and homeostasis, so it is not surprising that endothelial dysfunction and dysregulation results in a variety of different pathophysiological conditions. The large number of vascular-related disorders and the emergence of angiogenesis as a major hallmark of cancer has led to significant interest in the development of drugs that target the vasculature. While several in vivo models exist to study developmental and pathological states of blood vessels, few in vitro assays have been developed that capture the significant complexity of the vascular microenvironment. Here, we describe a high content endothelial colony forming cells (ECFC)/adipose-derived stem cell (ADSC) coculture assay that captures many elements of in vivo vascular biology and is ideal for in vitro screening of compounds for pro- or anti-angiogenic activities.

Keywords: Adipose-derived stem cell (ADSC); Angiogenesis; CD31; Cord formation; Endothelial cell; High content assay (HCA); Pericyte; Phenotypic drug discovery; Smooth muscle actin (SMA); Vascular endothelial growth factor (VEGF).

MeSH terms

  • Biological Assay*
  • Biomarkers
  • Cell Culture Techniques
  • Coculture Techniques*
  • Data Interpretation, Statistical
  • Dose-Response Relationship, Drug
  • Drug Discovery / methods
  • Endothelial Cells / cytology*
  • Endothelial Cells / metabolism
  • Image Processing, Computer-Assisted
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / metabolism
  • Microscopy
  • Neovascularization, Physiologic* / drug effects
  • Phenotype
  • Stem Cells / cytology
  • Stem Cells / metabolism
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • Biomarkers
  • Vascular Endothelial Growth Factor A