Ex vivo generation of functional dendritic cells from mobilized CD34+ hematopoietic stem cells.
10.3349/ymj.1998.39.4.328
- Author:
Yoo Hong MIN
1
;
Seung Tae LEE
;
Kyung Mi CHOI
;
So Young CHONG
;
Hyun Ok KIM
;
Jee Sook HAHN
;
Yun Woong KO
Author Information
1. Department of Internal Medicine, Yonsei University College of Medicine, Seoul, Korea.
- Publication Type:Original Article
- Keywords:
Dendritic cells;
CD34;
peripheral blood hematopoietic stem cells
- MeSH:
Antigens, CD34/analysis*;
Dendritic Cells/physiology*;
Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology;
HLA-DR Antigens/analysis;
Hematopoietic Stem Cells/physiology*;
Human;
Interleukin-4/pharmacology;
Tumor Necrosis Factor/pharmacology
- From:Yonsei Medical Journal
1998;39(4):328-338
- CountryRepublic of Korea
- Language:English
-
Abstract:
The ability to generate dendritic cells (DCs) in sizeable numbers has enormous implications for the development of clinically-effective antigen presentation procedures for cancer immunotherapy. We evaluated the generation of immunostimulatory DCs from peripheral blood CD34+ cells collected from healthy donors. CD34+ cells purified from leukapheresis product were seeded at 1 x 10(4) cells/mL in complete medium supplemented with GM-CSF, TNF alpha, IL-4, c-kit ligand, and flt3 ligand (FL). By day 14 of culture in the presence of GM-CSF + TNF alpha, the total cell number increased by 23.4 +/- 5.4-fold compared to the starting number of CD34+ cells. When the c-kit and FL were added to GM-CSF and TNF alpha, the cell number increased by 109.8 +/- 11.2-fold without affecting the immunophenotype of recovered cells. Flow cytometric analysis indicated that cells with the markers of mature dendritic cells, i.e., CD1a +CD14 -HLA-DR+, and CD80+CD86+HLA-DR+, constituted 49.0% +/- 7.5%, and 38.9% +/- 6.5%, respectively. This pattern of expression of surface antigen was unchanged whether the c-kit ligand and/or FL was added. The irradiated CD1a+HLA-DR+ cells recovered from in vitro cultures elicit a vigorous proliferation of allogeneic peripheral blood T-cells, irrespective of cytokine combinations. These findings provide advantageous tools for the large-scale generation of DCs that are potentially usable for clinical protocols of immunotherapy or vaccination in patients undergoing cancer treatment.
