title: Bone marrow stromal cells prepared using AB serum and bFGF for hematopoietic stem cells expansion.
abstract: BACKGROUND
  An ex vivo culture system was previously established for stem cell expansion using human marrow stromal cells and serum-free medium. However, the stromal cells were prepared using long-term culture medium containing horse serum and FCS, which may transmit infectious diseases of xenogeneic origin. In this study, therefore, a method was established to prepare stromal cells using an AB serum-based medium. In the case that serum from a transplant recipient or PBPC donor is available, additional infectious diseases would not be transmitted.
  
  
  METHODS
  Cord blood CD34+ cells were cultured with thrombopoietin, stem cell factor, and flt3/flk2 ligand on a monolayer of human marrow primary stromal cells prepared using long-term culture medium or AB serum-based medium. After 2 weeks, clonogenic progenitor activity and SCID mouse-reconstituting cell activity were assayed. mRNA expression of cytokines and Notch ligand by stromal cells was also examined.
  
  
  RESULTS
  There were no remarkable differences in expansion-supporting activity and mRNA expression between stromal cells established by the two methods.
  
  
  CONCLUSIONS
  An ex vivo expansion system completely based on AB serum has been established.
authors.affiliation: Hokkaido Red Cross Blood Center, Sapporo, Japan. mikiyama@hokkaido.bc.jrc.or.jp
authors.forename: Miki
authors.initials: M
authors.lastname: Yamaguchi
authors.forename: Fumiya
authors.initials: F
authors.lastname: Hirayama
authors.forename: Shinobu
authors.initials: S
authors.lastname: Wakamoto
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Fujihara
authors.forename: Hideaki
authors.initials: H
authors.lastname: Murahashi
authors.forename: Norihiro
authors.initials: N
authors.lastname: Sato
authors.forename: Kenji
authors.initials: K
authors.lastname: Ikebuchi
authors.forename: Ken-ichi
authors.initials: K
authors.lastname: Sawada
authors.forename: Takao
authors.initials: T
authors.lastname: Koike
authors.forename: Mikinori
authors.initials: M
authors.lastname: Kuwabara
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Azuma
authors.forename: Hisami
authors.initials: H
authors.lastname: Ikeda
mesh_terms: D000017:ABO Blood-Group System / Q000097:blood / Q000494:pharmacology*; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D018929:Cell Culture Techniques / Q000379:methods / Q000592:standards; D002455:Cell Division / Q000187:drug effects; D018920:Coculture Techniques; D003114:Colony-Forming Units Assay; D005312:Fetal Blood / Q000166:cytology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D006133:Growth Substances / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000276:immunology; D006801:Humans; D016130:Immunophenotyping; D017053:Infection Control / Q000379:methods; D051379:Mice; D016513:Mice, SCID; D033581:Stem Cell Transplantation / Q000379:methods; D017154:Stromal Cells / Q000166:cytology*; D014183:Transplantation, Heterologous / Q000379:methods
chemical_list: D000017:ABO Blood-Group System; D006133:Growth Substances; D016222:Fibroblast Growth Factor 2