Extensive and long-term ex vivo production of dendritic cells from CD34 positive umbilical cord blood or bone marrow cells by novel culture system using mouse stroma.
We previously developed a system using murine strome (HESS-5), which could expand umbilical cord blood (UCB) stem and progenitor cells, especially CD34+/38- cells, in the presence of human recombinant cytokines. In this study, the ability of expanded UCB- or bone marrow (BM)-CD34+ cells to differentiate into dendritic cells (DCs) was examined. DCs could be induced either from short or long term cultured CD34+ cells after switching the cytokines from Flk-2/Flt-3 ligand, stem cell factor (SCF), thrombopoietin (TPO) to granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) (immature type) plus tumor necrosis factor (TNF)-alpha with stimulation by CD40L transfectant (mature type). Each immature or mature UCB-DCs showed a dextran uptake or a potent allo-T lymphocytes proliferative ability, respectively. Furthermore, those DCs from BM significantly stimulated auto-T lymphocytes in an antigen (varicella zoster virus) specific manner. In conclusion, a novel culture system using HESS-5 is useful to support a rapid and sustained generation of primitive myeloid cells which can develop into functional DCs.
Hagihara
M
M
Department of Hematology and Rheumatology, Tokai University School of Medicine, Bohseidai, Kanagawa, 259-1193, Isehara, Japan. masaoha@.is.icc.u-tokai.ac.jp
Li
C
C
Gansuvd
B
B
Munkhbat
B
B
Inoue
H
H
Shimakura
Y
Y
Tsuchiya
T
T
Ueda
Y
Y
Oki
M
M
Ando
K
K
Kato
S
S
Hotta
T
T
D000818:Animals; D018952:Antigens, CD34 / Q000032:analysis*; D000956:Antigens, Viral / Q000276:immunology; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002455:Cell Division; D002460:Cell Line; D016207:Cytokines / Q000494:pharmacology; D003713:Dendritic Cells / Q000276:immunology*; D003911:Dextrans / Q000378:metabolism; D005312:Fetal Blood / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D007519:Isoantigens / Q000276:immunology; D007700:Kinetics; D051379:Mice; D010641:Phenotype; D017154:Stromal Cells / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology
D018952:Antigens, CD34; D000956:Antigens, Viral; D016207:Cytokines; D003911:Dextrans; D007519:Isoantigens