Phenotype and function of GM-CSF independent dendritic cells generated by long-term propagation of rat bone marrow cells.
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.
Department of Research Surgery, National Center for Child Health and Development, Tokyo, Japan.
Hua
H
Yan
Tohko
T
Miyagi
Eigo
E
Satoh
Wataru
W
Sugiura
Naoki
N
Yamamoto
Hiromitsu
H
Kimura
D000818:Animals; D000954:Antigens, Surface / Q000276:immunology; D001854:Bone Marrow Cells / Q000166:cytology / Q000276:immunology*; D018929:Cell Culture Techniques / Q000379:methods*; D003433:Crosses, Genetic; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000276:immunology / Q000494:pharmacology; D006801:Humans; D016130:Immunophenotyping; D015850:Interleukin-6 / Q000276:immunology / Q000494:pharmacology*; D037181:Lectins, C-Type / Q000276:immunology; D007959:Lymphocyte Culture Test, Mixed; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D055651:NK Cell Lectin-Like Receptor Subfamily B; D011518:Proto-Oncogene Proteins / Q000276:immunology / Q000494:pharmacology*; D051381:Rats; D011919:Rats, Inbred Strains; D020794:Receptor Protein-Tyrosine Kinases / Q000276:immunology / Q000494:pharmacology*; D016038:Skin Transplantation / Q000276:immunology; D019089:Stem Cell Factor / Q000276:immunology / Q000494:pharmacology; D051941:fms-Like Tyrosine Kinase 3
D000954:Antigens, Surface; D015850:Interleukin-6; C527203:KLRB1 protein, human; D037181:Lectins, C-Type; D055651:NK Cell Lectin-Like Receptor Subfamily B; D011518:Proto-Oncogene Proteins; D019089:Stem Cell Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; C497970:FLT3 protein, human; C497971:Flt3 protein, mouse; D020794:Receptor Protein-Tyrosine Kinases; D051941:fms-Like Tyrosine Kinase 3