title: Comparison of monocyte enrichment by immuno-magnetic depletion or adherence for the clinical-scale generation of DC.
abstract: 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.
authors.affiliation: Nexell Therapeutics, Inc., Cellular Biology, 9 Parker, Irvine, CA 92618, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Suen
authors.forename: S M
authors.initials: SM
authors.lastname: Lee
authors.forename: F
authors.initials: F
authors.lastname: Aono
authors.forename: S
authors.initials: S
authors.lastname: Hou
authors.forename: M
authors.initials: M
authors.lastname: Loudovaris
authors.forename: G
authors.initials: G
authors.lastname: Ofstein
authors.forename: J G
authors.initials: JG
authors.lastname: Bender
mesh_terms: 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
chemical_list: D018941:Antigens, CD19; D018801:CD2 Antigens; D003911:Dextrans