Phenotype and function of GM-CSF independent dendritic cells generated by long-term propagation of rat bone marrow cells

Cell Immunol. 2004 Jun;229(2):117-29. doi: 10.1016/j.cellimm.2004.07.006.

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

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Surface / immunology
  • Bone Marrow Cells / cytology
  • Bone Marrow Cells / immunology*
  • Cell Culture Techniques / methods*
  • Crosses, Genetic
  • Dendritic Cells / cytology
  • Dendritic Cells / immunology*
  • Female
  • Flow Cytometry
  • Granulocyte-Macrophage Colony-Stimulating Factor / immunology
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
  • Humans
  • Immunophenotyping
  • Interleukin-6 / immunology
  • Interleukin-6 / pharmacology*
  • Lectins, C-Type / immunology
  • Lymphocyte Culture Test, Mixed
  • Male
  • Mice
  • Mice, Inbred C57BL
  • NK Cell Lectin-Like Receptor Subfamily B
  • Proto-Oncogene Proteins / immunology
  • Proto-Oncogene Proteins / pharmacology*
  • Rats
  • Rats, Inbred Strains
  • Receptor Protein-Tyrosine Kinases / immunology
  • Receptor Protein-Tyrosine Kinases / pharmacology*
  • Skin Transplantation / immunology
  • Stem Cell Factor / immunology
  • Stem Cell Factor / pharmacology
  • fms-Like Tyrosine Kinase 3

Substances

  • Antigens, Surface
  • Interleukin-6
  • KLRB1 protein, human
  • Lectins, C-Type
  • NK Cell Lectin-Like Receptor Subfamily B
  • Proto-Oncogene Proteins
  • Stem Cell Factor
  • Granulocyte-Macrophage Colony-Stimulating Factor
  • FLT3 protein, human
  • Flt3 protein, mouse
  • Receptor Protein-Tyrosine Kinases
  • fms-Like Tyrosine Kinase 3