1. Progressive and controlled development of mouse dendritic cells from Flt3+CD11b+ progenitors in vitro.
- Author
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Hieronymus T, Gust TC, Kirsch RD, Jorgas T, Blendinger G, Goncharenko M, Supplitt K, Rose-John S, Müller AM, and Zenke M
- Subjects
- Adoptive Transfer, Amino Acid Sequence, Animals, Antigens, Surface biosynthesis, Antigens, Surface genetics, Bone Marrow Cells cytology, Bone Marrow Cells immunology, Bone Marrow Cells metabolism, Cells, Cultured, Chemotaxis, Leukocyte immunology, Cytokines biosynthesis, Dendritic Cells immunology, Dendritic Cells metabolism, Epitopes, T-Lymphocyte administration & dosage, Epitopes, T-Lymphocyte immunology, Gene Expression Profiling, Hematopoietic Stem Cells immunology, Hematopoietic Stem Cells metabolism, Immunophenotyping, Lymphocyte Activation, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Mice, Transgenic, Molecular Sequence Data, T-Lymphocytes immunology, T-Lymphocytes metabolism, T-Lymphocytes transplantation, fms-Like Tyrosine Kinase 3, CD11b Antigen biosynthesis, Cell Differentiation immunology, Dendritic Cells cytology, Hematopoietic Stem Cells cytology, Proto-Oncogene Proteins biosynthesis, Receptor Protein-Tyrosine Kinases biosynthesis
- Abstract
Dendritic cells (DC) represent key regulators of the immune system, yet their development from hemopoietic precursors is poorly defined. In this study, we describe an in vitro system for amplification of a Flt3(+)CD11b(+) progenitor from mouse bone marrow with specific cytokines. Such progenitor cells develop into both CD11b(+) and CD11b(-) DC, and CD8alpha(+) and CD8alpha(-) DC in vivo. Furthermore, with GM-CSF, these progenitors synchronously differentiated into fully functional DC in vitro. This two-step culture system yields homogeneous populations of Flt3(+)CD11b(+) progenitor cells in high numbers and allows monitoring the consecutive steps of DC development in vitro under well-defined conditions. We used phenotypic and functional markers and transcriptional profiling by DNA microarrays to study the Flt3(+)CD11b(+) progenitor and differentiated DC. We report here on an extensive analysis of the surface Ag expression of Flt3(+)CD11b(+) progenitor cells and relate that to surface Ag expression of hemopoietic stem cells. Flt3(+)CD11b(+) progenitors studied exhibit a broad overlap of surface Ags with stem cells and express several stem cell Ags such as Flt3, IL-6R, c-kit/SCF receptor, and CD93/AA4.1, CD133/AC133, and CD49f/integrin alpha(6). Thus, Flt3(+)CD11b(+) progenitors express several stem cell surface Ags and develop into both CD11b(+) and CD11b(-) DC, and CD8alpha(+) and CD8alpha(-) DC in vivo, and thus into both of the main conventional DC subtypes.
- Published
- 2005
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