Abstract
GM-CSF is believed to be an essential factor for growth and differentiation of myeloid dendritic cells (DC). Employing a low-density fraction of rat bone marrow cells, we attempted to generate DC with human Flt-3/Flk-2 and IL-6. In this culture system, typical DC gradually appeared without exogenous GM-CSF supplement. Phenotypes and functions of the DC were examined. Evidence provided that the most efficient long-term outgrowth of DC progenitors was obtained by GM-CSF independent culture systems with the aid of Flt3/Flk-2 and IL-6, not with c-kit ligand and IL-6. Furthermore, CD103 (OX-62), which is widely used for rat DC separation, was found to be insufficient for enriching DC, due to the down-regulation of the marker. However, the most efficient selection of rat DC was made by CD161a (NKR-P1A), a C-type lectin family. The GM-CSF independent DC was functionally active in vitro as well as in vivo assays.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antigens, Surface / immunology
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Bone Marrow Cells / cytology
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Bone Marrow Cells / immunology*
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Cell Culture Techniques / methods*
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Crosses, Genetic
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Dendritic Cells / cytology
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Dendritic Cells / immunology*
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Female
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Flow Cytometry
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Granulocyte-Macrophage Colony-Stimulating Factor / immunology
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Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
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Humans
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Immunophenotyping
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Interleukin-6 / immunology
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Interleukin-6 / pharmacology*
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Lectins, C-Type / immunology
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Lymphocyte Culture Test, Mixed
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Male
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Mice
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Mice, Inbred C57BL
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NK Cell Lectin-Like Receptor Subfamily B
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Proto-Oncogene Proteins / immunology
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Proto-Oncogene Proteins / pharmacology*
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Rats
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Rats, Inbred Strains
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Receptor Protein-Tyrosine Kinases / immunology
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Receptor Protein-Tyrosine Kinases / pharmacology*
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Skin Transplantation / immunology
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Stem Cell Factor / immunology
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Stem Cell Factor / pharmacology
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fms-Like Tyrosine Kinase 3
Substances
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Antigens, Surface
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Interleukin-6
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KLRB1 protein, human
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Lectins, C-Type
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NK Cell Lectin-Like Receptor Subfamily B
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Proto-Oncogene Proteins
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Stem Cell Factor
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Granulocyte-Macrophage Colony-Stimulating Factor
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FLT3 protein, human
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Flt3 protein, mouse
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Receptor Protein-Tyrosine Kinases
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fms-Like Tyrosine Kinase 3