Wnt3 modulates the characteristics and cobblestone area-supporting activity of human stromal cells

Exp Hematol. 2004 Dec;32(12):1194-203. doi: 10.1016/j.exphem.2004.08.010.


Objective: Our objective was to investigate the expression and significance of Wnt proteins in adult human hematopoietic-supporting stromal cells.

Methods: Degenerate reverse transcription-polymerase chain reaction was performed to screen telomerized human stromal cells (hTERT-stromal cells) and multipotent mesenchymal cells (hTERT-MSCs) for expression of Wnt genes. We studied the actions of Wnt proteins by overexpressing them in stromal cells and MSCs by retrovirus-mediated gene transfer.

Results: The hTERT-stromal and primary stromal cells expressed Wnt5A, while hTERT-MSCs and primary MSCs expressed Wnt3 and Wnt5A. Gene transfer of Wnt5A slightly reduced the growth rate of hTERT-stromal cells, but did not affect their morphology. In contrast, gene transfer of Wnt3 into both hTERT-stromal cells and hTERT-MSCs enhanced Wnt-betacatenin signaling, and caused remarkable morphological changes and growth retardation. Upon 2-week co-culture, expansion of clonogenic cells on Wnt5A-stromal cells was superior to that on control stromal cells. However, expansion of CD34+ cells on Wnt3-stromal cells did not differ from that on control stromal cells. Moreover, there was a drastic reduction in the formation of cobblestone area (CA) underneath Wnt3-stromal cells compared with that underneath control stromal cells.

Conclusion: These results suggest that Wnt3 plays an important role in regulating characteristics and CA support activity of stromal cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD34 / metabolism
  • Cell Differentiation / genetics
  • Cell Differentiation / physiology
  • Cell Growth Processes / genetics
  • Cell Growth Processes / physiology
  • Coculture Techniques
  • Gene Expression Regulation / genetics
  • Gene Expression Regulation / physiology
  • Hematopoietic Stem Cells / cytology*
  • Hematopoietic Stem Cells / physiology
  • Humans
  • Mesoderm / cytology*
  • Mesoderm / physiology
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / physiology
  • Proteins / genetics
  • Proteins / metabolism*
  • Proto-Oncogene Proteins / genetics
  • Proto-Oncogene Proteins / metabolism
  • Stromal Cells / cytology
  • Stromal Cells / physiology*
  • Telomerase / genetics
  • Telomerase / metabolism
  • Transduction, Genetic
  • Wnt Proteins
  • Wnt-5a Protein
  • Wnt3 Protein


  • Antigens, CD34
  • Proteins
  • Proto-Oncogene Proteins
  • WNT3 protein, human
  • WNT5A protein, human
  • Wnt Proteins
  • Wnt-5a Protein
  • Wnt3 Protein
  • Telomerase