Optimizing preparation of normal dendritic cells and bcr-abl+ mature dendritic cells derived from immunomagnetically purified CD14+ cells

J Hematother Stem Cell Res. 2000 Feb;9(1):95-101. doi: 10.1089/152581600319676.


The goal of this work was to optimize dendritic cell (DC) preparations obtained from patients suffering from chronic myeloid leukemia (CML) and compare them with DC prepared from normal CD14+ mononuclear cells (MNC). We studied normal DC and bcr-abl+ leukemic DC (CML-DC) yields, expression of membrane molecules, differentiation status, and ability to stimulate T cells. We isolated DC precursors from PBMC by CD14-specific immunoadsorption and cultured them for 7 days in GM-CSF and IL-4, followed by a 3-day incubation to fully differentiate the cells. We evaluated cultures of CML-DC using RPMI 1640 medium supplemented with FBS and X-VIVO 15 medium containing human AB serum. In contrast to cells matured in RPMI 1640, virtually all cells incubated in X-VIVO 15 expressed CD83, a marker of mature DC. CML-DC and normal DC were indistinguishable in expression of CD83, resulting in the highest percentage reported so far. The yields of normal DC and CML-DC from CD14+ cells were indistinguishable. The percentage of bcr-abl+ cells in PBMC varied among patients between 65% and 97% and the final CML-DC preparations were >98% bcr-abl+ the highest purity of bcr-abl+ cells to date. Normal DC and CML-DC were equally effective in stimulating proliferation of allogeneic and autologous T cells. These techniques provide highly enriched, mature, functional CML-DC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Division
  • Coculture Techniques
  • Culture Media
  • Culture Media, Serum-Free
  • Cytokines
  • Dendritic Cells / cytology*
  • Dendritic Cells / drug effects
  • Fusion Proteins, bcr-abl / blood*
  • Humans
  • Immunomagnetic Separation
  • In Situ Hybridization, Fluorescence
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / blood
  • Leukocytes, Mononuclear
  • Lipopolysaccharide Receptors / blood*
  • Specimen Handling
  • T-Lymphocytes / cytology


  • Culture Media
  • Culture Media, Serum-Free
  • Cytokines
  • Lipopolysaccharide Receptors
  • Fusion Proteins, bcr-abl