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


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.


CONCLUSIONS
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.
Rouard
Helene
H
Cell Therapy Unit of EFS Ile de France-site Henri Mondor, 51 Avenue Marechal De Lattre de Tassigny, 94010 Creteil Cedex, France. helene.rouard@hmn.ap-hop-paris.fr
Leon
Anne
A
De Reys
Stef
S
Taylor
Linda
L
Logan
Josephine
J
Marquet
Jeanine
J
Jouault
Helene
H
Loper
Kathy
K
Maison
Patrick
P
Delfau-Larue
Marie-Helene
MH
Beaujean
Françoise
F
Farcet
Jean-Pierre
JP
Noga
Stephen J
SJ
D018941:Antigens, CD19 / Q000032:analysis; D001781:Blood Component Removal; D001792:Blood Platelets; D017252:CD3 Complex / Q000032:analysis; D019002:CD56 Antigen / Q000032:analysis; D002452:Cell Count; D002454:Cell Differentiation; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D048429:Cell Size; D003713:Dendritic Cells / Q000166:cytology*; D005434:Flow Cytometry; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D016130:Immunophenotyping; D007167:Immunotherapy; D015847:Interleukin-4 / Q000494:pharmacology; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D009000:Monocytes / Q000166:cytology*
D018941:Antigens, CD19; D017252:CD3 Complex; D019002:CD56 Antigen; D018950:Lipopolysaccharide Receptors; D015847:Interleukin-4; D016178:Granulocyte-Macrophage Colony-Stimulating Factor