Comparison of monocyte enrichment by immuno-magnetic depletion or adherence for the clinical-scale generation of DC

• Published in: Cytotherapy (Oxford, England) Vol. 3; no. 5; pp. 365 - 375
• Main Authors: Suen, Y., Lee, S.-M., Aono, F., Hou, S., Loudovaris, M., Ofstein, G., Bender, J.G.
• Format: Journal Article
• Language: English
• Published: England Elsevier Inc 01.09.2001; Informa UK Ltd
• Subjects: Antigens, CD19 - immunology; B-Lymphocytes - cytology; B-Lymphocytes - immunology; CD2 Antigens - immunology; Cell Adhesion; Cell Culture Techniques - instrumentation; Cell Culture Techniques - methods; Cell Division; Cell Separation - instrumentation; Cell Separation - methods; Cell Survival; clinical scale; Dendritic cell; Dendritic Cells - cytology; Dendritic Cells - immunology; Dextrans - metabolism; Flow Cytometry; Humans; Immunomagnetic Separation - instrumentation; Immunomagnetic Separation - methods; Immunophenotyping; immunotherapy; Isolex 300i magnetic cell selector; Monocytes - cytology; Pinocytosis; SteriCell culture bags; T-Lymphocytes - cytology; T-Lymphocytes - immunology; Dendritic cell; clinical scale; SteriCell culture bags; immunotherapy; Isolex 300i magnetic cell selector
• ISSN: 1465-3249; 1477-2566
• DOI: 10.1080/146532401753277184

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. 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. ISO-DC and AD-DC from SteriCell bags showed similar viability. Higher purity of ISO-DC than AD-DC was measured by forwardand 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. These data indicated that functional DC were generated in gas permeable SteriCell culture bags from both ISO- and AD-monocytes in a closed system.