title: Wnt3 modulates the characteristics and cobblestone area-supporting activity of human stromal cells.
abstract: 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.
  
  
  CONCLUSIONS
  These results suggest that Wnt3 plays an important role in regulating characteristics and CA support activity of stromal cells.
authors.affiliation: Forth Department of Internal Medicine, Sapporo Medical University School of Medicine, Sapporo, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Chiba
authors.forename: Masayoshi
authors.initials: M
authors.lastname: Kobune
authors.forename: Junji
authors.initials: J
authors.lastname: Kato
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Kawano
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Ito
authors.forename: Kiminori
authors.initials: K
authors.lastname: Nakamura
authors.forename: Sumiyo
authors.initials: S
authors.lastname: Asakura
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Hamada
authors.forename: Yoshiro
authors.initials: Y
authors.lastname: Niitsu
mesh_terms: D018952:Antigens, CD34 / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology; D048708:Cell Growth Processes / Q000235:genetics / Q000502:physiology; D018920:Coculture Techniques; D005786:Gene Expression Regulation / Q000235:genetics / Q000502:physiology; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000502:physiology; D006801:Humans; D008648:Mesoderm / Q000166:cytology* / Q000502:physiology; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology*; D019098:Telomerase / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; D060508:Wnt3 Protein
chemical_list: D018952:Antigens, CD34; D011506:Proteins; D011518:Proto-Oncogene Proteins; C555507:WNT3 protein, human; C085389:WNT5A protein, human; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; D060508:Wnt3 Protein; D019098:Telomerase