Comparison of monocyte enrichment by immuno-magnetic depletion or adherence for the clinical-scale generation of DC.
BACKGROUND
DC generated from monocytes have been used for vaccines. We have developed a monocyte enrichment procedure by depleting T and B cells with anti-CD2 and anti-CD19 Abs using the automated Isolex 300i magnetic cell selector for clinical-scale DC generation in gas permeable SteriCell culture bags. We have also compared DC function, yield and purity of DC generated from adherent monocytes using culture bags in a closed system, with DC generated in conventional tissue culture flasks.


METHODS
Monocytes were enriched from normal donor apheresis products using CD2/19 depletion with experimental software on the Isolex 300i (ISO), adherence (AD) to SteriCell bags and to T175 flasks and then cultured for 7 days in serum-free X-VIVO 15 media with GM-CSF and IL-4. Phenotype and dextran uptake were analyzed by flow cytometry and allogeneic MLR was also evaluated.


RESULTS
ISO-DC and AD-DC from SteriCell bags showed similar viability. Higher purity of ISO-DC than AD-DC was measured by forward- and side-scatter flow cytometry. Similar expression of CD1a, CD80, CD86 and CD83 were observed in both ISO-DC and AD-DC. Similar dextran uptake and allo MLR were also observed.


CONCLUSIONS
These data indicated that functional DC were generated in gas permeable SteriCell culture bags from both ISO- and AD-monocytes in a closed system.
Suen
Y
Y
Nexell Therapeutics, Inc., Cellular Biology, 9 Parker, Irvine, CA 92618, USA.
Lee
S M
SM
Aono
F
F
Hou
S
S
Loudovaris
M
M
Ofstein
G
G
Bender
J G
JG
D018941:Antigens, CD19 / Q000276:immunology; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology; D018801:CD2 Antigens / Q000276:immunology; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D002455:Cell Division; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D002470:Cell Survival; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology; D003911:Dextrans / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D018189:Immunomagnetic Separation / Q000295:instrumentation* / Q000379:methods*; D016130:Immunophenotyping; D009000:Monocytes / Q000166:cytology*; D010873:Pinocytosis; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology
D018941:Antigens, CD19; D018801:CD2 Antigens; D003911:Dextrans