Endothelial differentiation of mesenchymal stromal cells

PLoS One. 2012;7(10):e46842. doi: 10.1371/journal.pone.0046842. Epub 2012 Oct 4.

Abstract

Human mesenchymal stromal cells (hMSCs) are increasingly used in regenerative medicine for restoring worn-out or damaged tissue. Newly engineered tissues need to be properly vascularized and current candidates for in vitro tissue pre-vascularization are endothelial cells and endothelial progenitor cells. However, their use in therapy is hampered by their limited expansion capacity and lack of autologous sources. Our approach to engineering large grafts is to use hMSCs both as a source of cells for regeneration of targeted tissue and at the same time as the source of endothelial cells. Here we investigate how different stimuli influence endothelial differentiation of hMSCs. Although growth supplements together with shear force were not sufficient to differentiate hMSCs with respect to expression of endothelial markers such as CD31 and KDR, these conditions did prime the cells to differentiate into cells with an endothelial gene expression profile and morphology when seeded on Matrigel. In addition, we show that endothelial-like hMSCs are able to create a capillary network in 3D culture both in vitro and in vivo conditions. The expansion phase in the presence of growth supplements was crucial for the stability of the capillaries formed in vitro. To conclude, we established a robust protocol for endothelial differentiation of hMSCs, including an immortalized MSC line (iMSCs) which allows for reproducible in vitro analysis in further studies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers / metabolism
  • Bone Marrow Cells / cytology
  • Cell Differentiation* / drug effects
  • Cell Line
  • Cell Proliferation / drug effects
  • Cell Separation
  • Cell Shape / drug effects
  • Endothelial Cells / cytology*
  • Endothelial Cells / drug effects
  • Endothelial Cells / metabolism
  • Humans
  • Lactic Acid / chemistry
  • Lactic Acid / pharmacology
  • Male
  • Mesenchymal Stem Cells / cytology*
  • Mice
  • Polyesters
  • Polyglycolic Acid / chemistry
  • Polyglycolic Acid / pharmacology
  • Polylactic Acid-Polyglycolic Acid Copolymer
  • Polymers / chemistry
  • Tissue Engineering
  • Tissue Scaffolds / chemistry
  • Wound Healing / drug effects

Substances

  • Biomarkers
  • Polyesters
  • Polymers
  • Polylactic Acid-Polyglycolic Acid Copolymer
  • Polyglycolic Acid
  • Lactic Acid
  • poly(lactide)

Grant support

This work was sponsored by a research grant from De Stichting voor de Technische Wetenschappen (STW) to JdB. The authors also acknowledge financial support from the Netherlands Institute of Regenerative Medicine. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.