Synergistic effects of hepatocyte growth factor on human cord blood CD34+ progenitor cells are the result of c-met receptor expression

Stem Cells. 1996 Sep;14(5):592-602. doi: 10.1002/stem.140592.

Abstract

Hepatocyte growth factor (HGF) is a pleiotropic growth factor which, in addition to its mitogenic potency for primary hepatocytes, also has a role in the regulation of cell motility, cell growth and morphogenesis. In the present study, we show that c-met, the high-affinity receptor for HGF, is expressed on human cord blood (CB) CD34+ progenitor cells and CD34+Thy-1+ Lin-(lin-) cells. We have investigated the capacity of HGF to synergize with other growth factors to induce colony formation by CB CD34+ progenitor cells. CD34+ cells were cultured in semisolid medium containing serum with increasing concentrations of GM-CSF, G-CSF, macrophage colony-stimulating factor (M-CSF), stem cell factor (SCF), interleukin 3 (IL-3) and IL-11 alone or in combination with HGF. HGF acted as a potent synergist and enhanced, up to fourfold, colony formation induced by GM-CSF, G-CSF or M-CSF. HGF in combination with SCF, IL-3 or IL-11 did not induce proliferation of colony forming units-granulocyte macrophage (CFU-GM) above control levels. In serum-deprived cultures, HGF was only detectably synergistic with IL-11, and all other culture combinations showed no proliferation. To determine whether the stimulatory effect of IL-11 and the synergistic effect of HGF in the absence of serum could be attributed to the effect of these two cytokines on stem cells, IL-11-stimulated and unstimulated lin- cells were analyzed for expression of c-met. CD34+Thy-1+Lin- cells were positive for c-met, both in the presence and absence of IL-11 stimulation, and Northern analysis indicated that c-met RNA expression was upregulated in response to IL-11 compared to unstimulated controls. These results provide strong evidence for upregulation of the HGF receptor on primitive hematopoietic cells by IL-11, and for the synergistic role of HGF in colony formation by hematopoietic stem cells.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antigens, CD34 / analysis
  • Blood Proteins / pharmacology
  • Blotting, Northern
  • Cells, Cultured / drug effects
  • Cells, Cultured / metabolism
  • Culture Media, Serum-Free / pharmacology
  • Drug Synergism
  • Female
  • Fetal Blood / cytology*
  • Flow Cytometry
  • Granulocyte Colony-Stimulating Factor / pharmacology
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
  • Hematopoiesis / drug effects
  • Hepatocyte Growth Factor / pharmacology*
  • Humans
  • Immunohistochemistry
  • Interleukin-11 / pharmacology
  • Macrophage Colony-Stimulating Factor / pharmacology
  • Proto-Oncogene Proteins c-met
  • RNA, Messenger / metabolism
  • Receptor Protein-Tyrosine Kinases / analysis
  • Receptor Protein-Tyrosine Kinases / biosynthesis*
  • Receptor Protein-Tyrosine Kinases / genetics
  • Stem Cells / chemistry
  • Stem Cells / drug effects*
  • Stem Cells / metabolism
  • Thy-1 Antigens / analysis
  • Up-Regulation / drug effects

Substances

  • Antigens, CD34
  • Blood Proteins
  • Culture Media, Serum-Free
  • Interleukin-11
  • RNA, Messenger
  • Thy-1 Antigens
  • Granulocyte Colony-Stimulating Factor
  • Hepatocyte Growth Factor
  • Macrophage Colony-Stimulating Factor
  • Granulocyte-Macrophage Colony-Stimulating Factor
  • Proto-Oncogene Proteins c-met
  • Receptor Protein-Tyrosine Kinases