A closed and single-use system for monocyte enrichment: potential for dendritic cell generation for clinical applications

Transfusion. 2003 Apr;43(4):481-7. doi: 10.1046/j.1537-2995.2003.00353.x.

Abstract

Background: This study evaluated the ability of a modified cell separator (Cobe Spectra Apheresis) system to isolate monocytes (MOs) by elutriation. The evaluation was performed in two independent international laboratories. The capacity of collected MOs to differentiate into dendritic cells (DCs) was also assessed.

Study design and methods: MNCs from platelet apheresis residues were elutriated on a modified cell separator (Cobe Spectra Apheresis system) using a custom disposable set. Cells were separated according to their size and density. Recovery and purity of the collected cell product were evaluated by impedance counting and flow cytometry. DCs were differentiated in culture from the elutriated MOs and characterized by their surface markers and stimulatory capacity in a mixed WBC reaction assay.

Results: Six apheresis mononuclear cell products were used by each laboratory. The separation was achieved in less than 1 hour. Collected MOs had the potential to differentiate into DCs.

Conclusion: The modified cell separator is an easy and fast device to obtain highly enriched MOs with a DC differentiation potential. The system is closed and employs a single-use disposable set and is more amenable to good tissue practice. This method could become a valuable tool for DC-based active immunotherapy.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD19 / analysis
  • Blood Component Removal
  • Blood Platelets
  • CD3 Complex / analysis
  • CD56 Antigen / analysis
  • Cell Count
  • Cell Differentiation
  • Cell Separation / instrumentation
  • Cell Separation / methods*
  • Cell Size
  • Dendritic Cells / cytology*
  • Flow Cytometry
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
  • Humans
  • Immunophenotyping
  • Immunotherapy
  • Interleukin-4 / pharmacology
  • Lipopolysaccharide Receptors / analysis
  • Monocytes / cytology*

Substances

  • Antigens, CD19
  • CD3 Complex
  • CD56 Antigen
  • Lipopolysaccharide Receptors
  • Interleukin-4
  • Granulocyte-Macrophage Colony-Stimulating Factor