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.
Abbreviations
- DMSCs
- Dermis-derived mesenchymal stem cells
- VECs
- Vascular endothelial cells
- VEGF
- Vascular endothelial growth factor
- ECs
- Endothelial cells
- EPCs
- Endothelial progenitor cells
- MSCs
- Mesenchymal stem cells
- BMSCs
- Bone marrow-derived MSCs
- bFGF
- Basic fibroblast growth factor
- BMP-4
- Bone morphogenetic protein-4
- HLA
- Human leucocyte antigen
- FITC
- Fluorescein isothiocyanate
- Dil-ac-LDL
- Dil-acetylated low-density lipoprotein
- ELISA
- Enzyme-linked immunosorbent assay
- IL-5
- Iinterleukin-5
- HGF
- Hepatocyte growth factor
- TGF-β1
- Transforming growth factor-β1
- GM-CSF
- Granulocyte-macrophage colony stimulating factor
- sVCAM-1
- Soluble vascular cell adhesion molecule1
- IGF-1
- Human insulin-like growth factor-1
- EGM-2
- Endothelial cell growth medium-2
- PDGF
- Platelet derived growth factor
- ET1
- Endothelin-1
- HAECs
- Human aortic endothelial cells.