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Production of large numbers of Langerhans' cells with intraepithelial migration ability in vitro
Author(s) -
Hubert Pascale,
Bousarghin Latifa,
Greimers Roland,
FranzenDetrooz Elizabeth,
Boniver Jacques,
Delvenne Philippe
Publication year - 2005
Publication title -
experimental dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.108
H-Index - 96
eISSN - 1600-0625
pISSN - 0906-6705
DOI - 10.1111/j.0906-6705.2005.00299.x
Subject(s) - birbeck granules , biology , langerhans cell , stem cell factor , microbiology and biotechnology , progenitor cell , immunology , haematopoiesis , cytokine , keratinocyte growth factor , stem cell , growth factor , immune system , receptor , biochemistry
Langerhans' cells (LCs) are a subset of immature dendritic cells (DCs) and play a key role in the initiation and regulation of immune responses. Functional studies of these cells have been hampered by difficulties in generating a large number of LCs in vitro . We describe a new method to efficiently generate immature DCs exhibiting morphological, immunohistochemical, and ultrastructural features of LCs (CD1a + , Birbeck Granules + , CD207 + , E‐cadherin + , cutaneous lymphocyte‐associated antigen + , and CCR6 + ) from a limited number of CD34 + cord blood progenitors. This method is based on a two‐step procedure consisting of an amplification phase followed by a terminal differentiation induction. The amplification step is initiated with a combination of hematopoietic growth factors (thrombopoietin/stem cell factor/fetal liver tyrosine kinase‐3 ligand), cytokines (granulocyte–macrophage colony‐stimulating factor, tumor necrosis factor‐α, and interleukin‐4), and 5 ng/ml of transforming growth factor (TGF)‐β1. The differentiation is induced by increasing the concentration of TGF‐β1 to 12.5 ng/ml. These culture conditions were efficient for generating a large number of immature LCs (8.74 × 10 6 ± 3.2) from 15 × 10 4 CD34 + progenitor cells. In addition, these LCs were shown to be able to infiltrate an in vitro reconstructed epithelium. Because LCs play an important role in the mucosal immunity, this technique could be useful to study their interactions with epithelial pathogenic agents and to perform pharmacological, toxicological, and clinical research.