Cell-Surface MMP-9 Protein Is a Novel Functional Marker to Identify and Separate Proangiogenic Cells from Early Endothelial Progenitor Cells Derived from CD133(+) Cells

Stem Cells. 2016 May;34(5):1251-62. doi: 10.1002/stem.2300. Epub 2016 Feb 22.

Abstract

To develop cell therapies for ischemic diseases, endothelial progenitor cells (EPCs) have been expected to play a pivotal role in vascular regeneration. It is desirable to use a molecular marker that is related to the function of the cells. Here, a quantitative polymerase chain reaction array revealed that early EPCs derived from CD133(+) cells exhibited significant expression of MMP-9. Some populations of early EPCs expressed MMP-9 on the cell surface and others did not. We also attempted to separate the proangiogenic fraction from early EPCs derived from CD133(+) cells using a functional cell surface marker, and we then analyzed the MMP-9(+) and MMP-9(-) cell fractions. The MMP-9(+) cells not only revealed higher invasion ability but also produced a high amount of IL-8. Moreover, the stimulative effect of MMP-9(+) cells on angiogenesis in vitro and in vivo was prohibited by anti-IL-8 antibody. These data indicate that MMP-9 is one of the useful cell surface markers for the separation of angiogenic cells. Our treatment of early EPCs with hyaluronidase caused not only a downregulation of cell-surface MMP-9 but also a decrease in invasion ability, indicating that membrane-bound MMP-9, which is one of the useful markers for early EPCs, plays an important role in angiogenesis. Stem Cells 2016;34:1251-1262.

Keywords: Angiogenesis; CD133; Cell therapy; Endothelial progenitor cells; Matrix metalloproteinases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AC133 Antigen / metabolism*
  • Animals
  • Biomarkers / metabolism
  • Cell Fractionation
  • Cell Membrane / enzymology*
  • Cell Separation*
  • Endothelial Progenitor Cells / cytology*
  • Endothelial Progenitor Cells / drug effects
  • Endothelial Progenitor Cells / metabolism*
  • Flow Cytometry
  • Gene Expression Profiling
  • Hindlimb / blood supply
  • Hindlimb / pathology
  • Human Umbilical Vein Endothelial Cells / drug effects
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Humans
  • Hyaluronoglucosaminidase / pharmacology
  • Interleukin-8 / metabolism
  • Ischemia / pathology
  • Matrix Metalloproteinase 2 / metabolism
  • Matrix Metalloproteinase 9 / metabolism*
  • Matrix Metalloproteinase Inhibitors / pharmacology
  • Mice
  • Neovascularization, Physiologic* / drug effects
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism

Substances

  • AC133 Antigen
  • Biomarkers
  • Interleukin-8
  • Matrix Metalloproteinase Inhibitors
  • RNA, Messenger
  • Hyaluronoglucosaminidase
  • Matrix Metalloproteinase 2
  • Matrix Metalloproteinase 9