Generation of dendritic cells using cell culture bags--description of a method and review of literature.
Anticancer immunotherapy using dendritic cell-based vaccines is a strategy aimed at the induction and maintenance of immune responses against cancer cells. Clinical applications of dendritic cells (DCs) require stringent adherence to Good Manufacturing Practice (GMP) methods and rigorous standardization of DC-based vaccine preparation. Recently, closed systems for DC culture have been developed with a goal to minimize the risk of contamination. Here, we compare the yield, immunophenotype, and functional properties of DCs generated in Lifecell X-Fold culture bags and in plastic wells, both from adherence-selected monocytes, and review the current literature on closed systems for DC generation. We found that both the overall yield and the yield of CD83+ cells in cell culture bags was lower than in the standard culture method. No statistically significant differences were observed in the expression of DC immunophenotypic markers. The capability of DCs cultured in bags and in wells to induce the proliferation of allogeneic mononuclear cells were equivalent. The performance of DCs in mixed lymphocyte reaction correlated significantly (p = 0.005) with the CD83 expression but not with the CD80, CD86, HLA-DR, CD1a, and CD1c expression. We conclude that the immunophenotype and stimulatory properties of DCs cultured in closed cell culture bags are similar to those generated by conventional method using cell culture wells.
Büchler
T
T
Department of Clinical Hematology, Brno, Czech Republic. tbuchler@fnbrno.cz
Kovárová
L
L
Musilová
R
R
Bourková
L
L
Ocadlíková
D
D
Buliková
A
A
Hanák
L
L
Michálek
J
J
Hájek
R
R
D018929:Cell Culture Techniques / Q000379:methods; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology*; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D016130:Immunophenotyping; D007167:Immunotherapy / Q000379:methods*; D007959:Lymphocyte Culture Test, Mixed; D014612:Vaccines / Q000276:immunology*
D014612:Vaccines