Uraemia disrupts the vascular niche in a 3D co-culture system of human mesenchymal stem cells and endothelial cells

Nephrol Dial Transplant. 2012 Jul;27(7):2693-702. doi: 10.1093/ndt/gfr656. Epub 2011 Dec 29.

Abstract

Background: Recent studies identified mesenchymal stem cells (MSC) as major players in vascular remodelling and the sub-endothelial compartment as the stem cell niche. The uraemic microenvironment predisposes to increased levels of reactive oxygen species, accelerated ageing of endothelial cells (EC) and vascular sclerosis.

Methods: We generated an in vitro model of a vascular niche consisting of a three-dimensional collagen I/III gel containing MSC and EC. We recapitulated a uraemic microenvironment by supplementing the medium with 20% pooled sera from either healthy or uraemic patients.

Results: Under healthy conditions, MSC/EC co-culture in collagen gels resulted in vessel-like tube formation. In contrast, uraemic serum-induced expression of extracellular matrix proteins (real-time reverse transcription-polymerase chain reaction, immunohistochemistry) that was accompanied by significant collagen contraction and a myofibroblastic phenotype of MSC. Although the uraemic culture conditions stimulated EC proliferation (cell counting, BrdU assay) and did not induce apoptosis (7-amino-actinomycin, annexin V FACS analysis), the tube formation was disrupted in spite of significantly enhanced vascular endothelial growth factor-A messenger RNA expression. The excessive matrix synthesis and remodelling by uraemia-exposed MSC/EC was reminiscent of remodelling processes observed in arteries of 12 dialysis patients (using arteries from 10 children and 10 age-matched non-dialysis patients as controls).

Conclusion: Our data indicate a potential role of the sub-endothelial niche and its major cell types EC and MSC in the remodelling process of the vascular wall in CKD.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Apoptosis
  • Blood Vessels / cytology*
  • Blood Vessels / metabolism
  • Blotting, Western
  • Case-Control Studies
  • Cell Proliferation
  • Child
  • Coculture Techniques
  • Collagen / genetics
  • Collagen / metabolism
  • Endothelial Cells / cytology*
  • Endothelial Cells / metabolism
  • Extracellular Matrix
  • Female
  • Fibronectins / genetics
  • Fibronectins / metabolism
  • GPI-Linked Proteins / genetics
  • GPI-Linked Proteins / metabolism
  • Human Umbilical Vein Endothelial Cells / cytology
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Humans
  • Immunoenzyme Techniques
  • Male
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / metabolism
  • Middle Aged
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism
  • Netrins
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stem Cell Niche / physiology*
  • Uremia / metabolism*
  • Uremia / pathology*
  • Vascular Endothelial Growth Factor A / genetics
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • Fibronectins
  • GPI-Linked Proteins
  • NTNG1 protein, human
  • Nerve Tissue Proteins
  • Netrins
  • RNA, Messenger
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A
  • Collagen