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Generation of functionally mature dendritic cells from the multipotential stem cell line FDCP‐mix
Author(s) -
Schroeder Timm,
Lange Claudia,
Strehl John,
Just Ursula
Publication year - 2000
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/j.1365-2141.2000.02421.x
Subject(s) - cd40 , antigen presenting cell , stem cell , cd86 , biology , dendritic cell , microbiology and biotechnology , cd80 , haematopoiesis , follicular dendritic cells , progenitor cell , antigen , interleukin 3 , major histocompatibility complex , immune system , immunology , t cell , cytotoxic t cell , biochemistry , in vitro
Dendritic cells (DCs) are crucial components of the immune system because of their unique ability as antigen‐presenting cells for the initiation of a primary immune response. DCs, macrophages (Ms) and granulocytes (Gs) are believed to originate from a common myeloid progenitor cell. However, little is known about the molecular mechanisms leading to DC sublineage commitment. To establish a cell system that allows the molecular and biochemical analysis of DC differentiation and activation, we used the murine non‐leukaemic, multipotential stem cell line FDCP‐mix. FDCP‐mix cells were cultured in various amounts of GM‐colony stimulating factor (CSF) and interleukin (IL)‐4 for up to 16 d and analysed for morphology, expression of CD34, c‐kit, Gr‐1, Mac‐1, CD40, MHC‐I, MHC‐II and co‐stimulatory molecules (CD80, CD86) using flow cytometry, and for their capacity to present foreign antigen to autologous T cells. Up to d 7, the majority of FDCP‐mix cells consisted of cells differentiating along the G and M lineage. Thereafter, the number of dendritic cells increased until d 13. Differentiation along the DC lineage vs. the G and M lineage was favoured when FDCP‐mix cells were cultured in high concentration GM‐CSF (500 U/ml) throughout the culture and IL‐4 from d 9 onwards. The dendritic cells generated from FDCP‐mix cells were large, non‐adherent cells with veiled processes and expressed MHC II, CD40, CD80 and CD86. After pulsing with a foreign antigen (keyhole limpet haemocyanin), FDCP‐mix‐derived dendritic cells stimulated [ 3 H]‐thymidine incorporation of naive T‐cells in an autologous mixed lymphocyte reaction (MLR). Our results show that functionally mature dendritic cells are generated from the multipotential stem cell line FDCP‐mix. This cell line thus provides the unique possibility of establishing multipotential transgenic cell lines capable of differentiation along the DC lineage. The experimental system described here should prove a valuable tool for studying DC differentiation and function.