Efficient differentiation of vascular endothelial cells from dermal-derived mesenchymal stem cells induced by endothelial cell lines conditioned medium

Acta Histochem. 2018 Nov;120(8):734-740. doi: 10.1016/j.acthis.2018.08.004. Epub 2018 Aug 22.

Abstract

Objective: To directionally-differentiate dermis-derived mesenchymal stem cells (DMSCs) into vascular endothelial cells (VECs) in vitro, providing an experimental basis for studies on the pathogenesis and treatment of vascular diseases.

Methods: After separation by adherent culture, VEC line supernatant, vascular endothelial growth factor (VEGF), bone morphogenetic protein-4 and hypoxia were used for the differentiation of VECs from DMSCs. The cell type was authenticated by flow cytometry, matrigel angiogenesis assay in vitro, and immunofluorescent staining during differentiation. The VEGF concentration was investigated by enzyme-linked immunosorbent assay.

Results: After 28 days of differentiation, the cell surface marker CD31 was significantly positive (80%-90%) by flow cytometry in the VEC line-conditioned culture, which was significantly higher than in the other groups. Differentiated DMSCs had the ability to ingest Dil-ac-LDL and vascularize in the conditioned culture, but not in the other groups. In the VEC line supernatant, the concentration of VEGF was very low. The VEGF concentration changed along with the differentiation into VECs in the medium of the conditioned culture group.

Conclusion: VEC line supernatant can induce the differentiation of DMSCs into VECs, possibly through the pathway except VEGF.

Keywords: Conditioned medium; In vitro differentiation; Mesenchymal stem cells; Vascular endothelial cells.

MeSH terms

  • Adult
  • Cell Culture Techniques
  • Cell Differentiation
  • Culture Media, Conditioned
  • Dermis / cytology*
  • Endothelial Cells / cytology*
  • Endothelium / cytology*
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Flow Cytometry
  • Humans
  • Male
  • Mesenchymal Stem Cells / cytology*
  • Platelet Endothelial Cell Adhesion Molecule-1 / metabolism
  • Vascular Endothelial Growth Factor A

Substances

  • Culture Media, Conditioned
  • Platelet Endothelial Cell Adhesion Molecule-1
  • Vascular Endothelial Growth Factor A