Efficient ex vivo generation of human dendritic cells from mobilized CD34+ peripheral blood progenitors.
We tried to efficiently generate human dendritic cells (DCs) from CD34+ peripheral blood hematopoietic progenitor cells mobilized by high-dose chemotherapy and subsequent administration of granulocyte colony-stimulating factor, using a liquid suspension culture system. Among various combinations, the combination of c-kit ligand, flt-3 ligand, c-mpl ligand (TPO), and interleukin (IL)-4 most potently generated the number of CD1a+CD14- DCs in cultures containing granulocyte-macrophage colony-stimulating factor (GM-CSF) and tumor necrosis factor alpha (TNF-alpha). The delayed addition of IL-4 on day 6 of culture gave rise to an additional increase in the yield of CD1a+CD14-DCs that were characterized by the expression of HLA-ABC, HLA-DR, CD80, CD86, and CD83. The majority of the sorted CD1a-CD14+ cells derived from 6-day culture of CD34+ cells gave rise to CD1a+CD14- DCs and CD1a-CD14+ macrophages on day 12 of culture in the presence and absence of IL-4, respectively. These findings suggest that IL-4 promotes the differentiation of CD1a- CD14+ cells derived from mobilized CD34+ peripheral blood hematopoietic progenitors to CD1a+ CD14- DCs. The majority of these DCs expressed CD68 but not the Langerhans-associated granule antigen, a finding that suggests they emerge through the monocyte differentiation pathway. The addition of TPO and IL-4 to cultures did not affect the potential of DCs to stimulate the primary allogeneic T-cell response. These findings demonstrated that the combination of c-kit ligand plus flt-3 ligand plus TPO with GM-CSF plus TNF-alpha, followed by IL-4, is useful for ex vivo generation of human DCs from mobilized CD34+ peripheral blood progenitors.
Ohishi
K
K
Second Department of Internal Medicine, Mie University School of Medicine, Tsu, Japan.
Katayama
N
N
Mitani
H
H
Araki
H
H
Masuya
M
M
Suzuki
H
H
Hoshino
N
N
Miyashita
H
H
Nishii
K
K
Kageyama
S
S
Minami
N
N
Shiku
H
H
D018949:Antigens, CD1 / Q000032:analysis; D018952:Antigens, CD34 / Q000097:blood; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D003713:Dendritic Cells / Q000166:cytology*; D004347:Drug Interactions; D006133:Growth Substances / Q000494:pharmacology; D019337:Hematologic Neoplasms / Q000628:therapy; D019650:Hematopoietic Stem Cell Mobilization; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D006801:Humans; D016130:Immunophenotyping; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D007959:Lymphocyte Culture Test, Mixed
D018949:Antigens, CD1; D018952:Antigens, CD34; C419393:CD1a antigen; D003470:Culture Media; D006133:Growth Substances; D018950:Lipopolysaccharide Receptors