Bone marrow stromal cells prepared using AB serum and bFGF for hematopoietic stem cells expansion.
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.
Yamaguchi
Miki
M
Hokkaido Red Cross Blood Center, Sapporo, Japan. mikiyama@hokkaido.bc.jrc.or.jp
Hirayama
Fumiya
F
Wakamoto
Shinobu
S
Fujihara
Mitsuhiro
M
Murahashi
Hideaki
H
Sato
Norihiro
N
Ikebuchi
Kenji
K
Sawada
Ken-ichi
K
Koike
Takao
T
Kuwabara
Mikinori
M
Azuma
Hiroshi
H
Ikeda
Hisami
H
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
D000017:ABO Blood-Group System; D006133:Growth Substances; D016222:Fibroblast Growth Factor 2