title: A human stromal-based serum-free culture system supports the ex vivo expansion/maintenance of bone marrow and cord blood hematopoietic stem/progenitor cells.
abstract: OBJECTIVE
  We investigated the role of human stromal layers (hu-ST) on the ex vivo expansion/maintenance of human hematopoietic stem/progenitor cells (HSC) from adult bone marrow (BM) and umbilical cord blood (CB).
  
  
  METHODS
  BM and CB CD34(+)-enriched cells were cultured in serum-free medium supplemented with SCF, bFGF, LIF, and Flt-3, in the presence or absence of stroma, and analyzed for proliferation, phenotype, and clonogenic potential.
  
  
  RESULTS
  Significant expansion of BM and CB CD34(+) and CD34(+)CD38(-) cells were achieved in the presence of hu-ST. The differentiative potential of both BM and CB CD34(+)-enriched cells cocultured with hu-ST was primarily shifted toward the myeloid lineage, while maintaining/expanding a CD7(+) population. Clonogenic analysis of the expanded cells showed increases in progenitors of the myeloid lineage, including colony-forming unit-granulocyte, macrophage (CFU-GM) and colony-forming unit-granulocyte, erythroid, macrophage, megakaryocyte (CFU-Mix) for both BM (stroma and stroma-free conditions) and CB cells in the presence of stroma.
  
  
  CONCLUSIONS
  These results indicate that adult hu-ST in the presence of appropriate cytokines can be used to efficiently expand/maintain myeloid and lymphoid cell populations from human BM and CB HSC.
authors.affiliation: Department of Animal Biotechnology, University of Nevada, Reno, 89557, USA.
authors.forename: Cláudia Lobato
authors.initials: CL
authors.lastname: da Silva
authors.forename: Raquel
authors.initials: R
authors.lastname: Gonçalves
authors.forename: Kirsten B
authors.initials: KB
authors.lastname: Crapnell
authors.forename: Joaquim M S
authors.initials: JM
authors.lastname: Cabral
authors.forename: Esmail D
authors.initials: ED
authors.lastname: Zanjani
authors.forename: Graça
authors.initials: G
authors.lastname: Almeida-Porada
mesh_terms: D015703:Antigens, CD / Q000032:analysis; D018952:Antigens, CD34 / Q000097:blood; D001854:Bone Marrow Cells / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods; D002455:Cell Division / Q000502:physiology*; D016895:Culture Media, Serum-Free; D005312:Fetal Blood / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D007231:Infant, Newborn; D007700:Kinetics; D017154:Stromal Cells / Q000166:cytology* / Q000502:physiology; D013997:Time Factors
chemical_list: D015703:Antigens, CD; D018952:Antigens, CD34; D016895:Culture Media, Serum-Free