Differentiation of CD45‑/CD31+ lung side population cells into endothelial and smooth muscle cells in vitro

Int J Mol Med. 2019 Mar;43(3):1128-1138. doi: 10.3892/ijmm.2019.4053. Epub 2019 Jan 8.

Abstract

Side population (SP) cells are a small subpopulation of cells found in many mammalian tissues and organs, identified by their capacity to efflux Hoechst 33342 dye. They are enriched for stem/progenitor cell activity. SP cells isolated from the adult mouse lung can be separated into a CD45+ subset (bone marrow‑derived) and a CD45‑ subset that can be subdivided into CD31‑ and CD31+ subpopulations. CD45‑/CD31‑ lung SP (LSP) cells are known to be mesenchymal stem cells. However, CD45‑/CD31+ LSP cells are not fully characterized. In the present study, it was found that CD45‑/CD31+ LSP cells were able to form colonies. Based on the expression of vascular endothelial growth factor receptor 2 (VEGFR2), these cells were separated into VEGFR2‑ and VEGFR2+ cells. The CD45‑/CD31+/VEGFR2‑ LSP cells expressed genes characteristic of smooth muscle and endothelial progenitors, and were able to differentiate into smooth muscle and endothelial cells in vitro. The CD45‑/CD31+/VEGFR2+ LSP cells expressed genes characteristic of endothelial progenitors and gave rise to endothelial cells, although not smooth muscle, in vitro. The data demonstrate that CD45‑/CD31+/VEGFR2‑ LSP cells differentiated into CD45‑/CD31+/VEGFR2+ LSP cells and then endothelial cells, indicating that CD45‑/CD31+/VEGFR2+ LSP cells are likely to be derived from CD45‑/CD31+/VEGFR2‑ LSP cells. Taken together, the results suggest that CD45‑/CD31+ LSP cells can be separated into CD45‑/CD31+/VEGFR2‑ LSP cells, which may be progenitors of endothelial and smooth muscle, whereas CD45‑/CD31+/VEGFR2+ LSP cells may serve as late commitment endothelial progenitors in the adult mouse lung.

MeSH terms

  • Animals
  • Biomarkers
  • Cell Differentiation* / genetics
  • Cell Line, Tumor
  • Cell Separation / methods
  • Cells, Cultured
  • Colony-Forming Units Assay
  • Endothelial Cells / cytology*
  • Endothelial Cells / metabolism*
  • Female
  • Fluorescent Antibody Technique
  • Gene Expression
  • Immunophenotyping
  • Leukocyte Common Antigens / genetics
  • Leukocyte Common Antigens / metabolism
  • Lung / cytology*
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / metabolism
  • Mice
  • Myocytes, Smooth Muscle / cytology*
  • Myocytes, Smooth Muscle / metabolism*
  • Platelet Endothelial Cell Adhesion Molecule-1 / genetics
  • Platelet Endothelial Cell Adhesion Molecule-1 / metabolism
  • Side-Population Cells / cytology*
  • Side-Population Cells / metabolism*

Substances

  • Biomarkers
  • Platelet Endothelial Cell Adhesion Molecule-1
  • Leukocyte Common Antigens