title: An ex vivo model of hematopoietic stem cell mobilization.
abstract: BACKGROUND
  Mobilization of hematopoietic stem cells to the circulation facilitates their collection, thereby providing a non-marrow source of these cells for transplantation. Hematopoietic cytokine administration induces mobilization for most, but not all, donors. Because the underlying biology of mobilization is not well understood, improving the process on a rational basis is difficult. The design of an in vitro mobilization model was pursued to facilitate investigations of the process.
  
  
  METHODS
  MS5 murine stromal cell line cells were grown to confluence on microporous transwell membranes. Murine femoral marrow plugs were placed on top of the prepared transwell membranes. The transwells were then seated in wells containing media and hematopoietic growth factors. Cells that were released from the marrow plugs over time and migrated through the stromal layer into the wells were assayed for stem cell/progenitor cell characteristics.
  
  
  RESULTS
  Few or no GM-CSF (progenitors) were found in wells containing media alone or media plus mobilizing cytokines after 24 h. After 120 h, the numbers of cells in the cytokine-containing wells increased, as did the numbers of CD34(+) cells. Cells in the wells at the time progenitor cells were most frequent were shown to include side population (SP) hematopoietic stem cells. After 120 h in the presence of cytokines, cells pooled from the wells were transplanted to lethally irradiated mice. Eighty per cent of the transplanted mice survived 30 days or more, demonstrating that radioprotective stem cells were present in the wells.
  
  
  CONCLUSIONS
  An ex vivo model has been designed that may aid investigations of the various steps of stem cell mobilization.
authors.affiliation: Department of Medicine, University of Nebraska Medical Center, Omaha, Nebraska 68198-7680, USA.
authors.forename: A
authors.initials: A
authors.lastname: Kessinger
authors.forename: B
authors.initials: B
authors.lastname: O'Kane Murphy
authors.forename: J D
authors.initials: JD
authors.lastname: Jackson
authors.forename: J G
authors.initials: JG
authors.lastname: Sharp
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000032:analysis; D002460:Cell Line; D018920:Coculture Techniques; D004921:Erythropoietin; D005260:Female; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D016179:Granulocyte Colony-Stimulating Factor; D019650:Hematopoietic Stem Cell Mobilization / Q000379:methods*; D018380:Hematopoietic Stem Cell Transplantation; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins; D013997:Time Factors; D019172:Transplantation Conditioning; D014916:Whole-Body Irradiation
chemical_list: D018952:Antigens, CD34; D005456:Fluorescent Dyes; D011994:Recombinant Proteins; D004921:Erythropoietin; D016179:Granulocyte Colony-Stimulating Factor