Document Detail


Comparison of monocyte enrichment by immuno-magnetic depletion or adherence for the clinical-scale generation of DC.
MedLine Citation:
PMID:  11953016     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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. DISCUSSION: 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:
Y Suen; S M Lee; F Aono; S Hou; M Loudovaris; G Ofstein; J G Bender
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Publication Detail:
Type:  Comparative Study; Journal Article    
Journal Detail:
Title:  Cytotherapy     Volume:  3     ISSN:  1465-3249     ISO Abbreviation:  Cytotherapy     Publication Date:  2001  
Date Detail:
Created Date:  2002-04-15     Completed Date:  2003-01-24     Revised Date:  2008-04-24    
Medline Journal Info:
Nlm Unique ID:  100895309     Medline TA:  Cytotherapy     Country:  England    
Other Details:
Languages:  eng     Pagination:  365-75     Citation Subset:  IM    
Affiliation:
Nexell Therapeutics, Inc., Cellular Biology, 9 Parker, Irvine, CA 92618, USA.
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MeSH Terms
Descriptor/Qualifier:
Antigens, CD19 / immunology
Antigens, CD2 / immunology
B-Lymphocytes / cytology,  immunology
Cell Adhesion
Cell Culture Techniques / instrumentation*,  methods*
Cell Division
Cell Separation / instrumentation,  methods*
Cell Survival
Dendritic Cells / cytology*,  immunology
Dextrans / metabolism
Flow Cytometry
Humans
Immunomagnetic Separation / instrumentation*,  methods*
Immunophenotyping
Monocytes / cytology*
Pinocytosis
T-Lymphocytes / cytology,  immunology
Chemical
Reg. No./Substance:
0/Antigens, CD19; 0/Antigens, CD2; 9004-54-0/Dextrans

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