Abstract
Background and Objectives: Dendritic cells (DCs), the most potent of antigen–presenting cells, can be generated in vitro from bone marrow or blood progenitor cells. We have developed a method for producing such cells from mobilised peripheral blood CD34+ cells in the absence of bovine products. Methods: The culture system developed used X–Vivo 10 culture medium with 10% autologous serum, rhGM–CSF, rhTNF–α and rhIL–4. Large–scale cultures were performed in Stericell 12 inch × 15 inch culture bags. Results: In 12–small–scale experiments, over 14 days, there was a median 38–fold increase in cell numbers of which 12.8% were DCs as defined by immunophenotyping. These cells had potent DC activity in functional assays. In two clinical–scale experiments, a 5.7– and 10–fold expansion of total cell numbers was obtained, with 8.2 and 18% of the final population being DCs, respectively. Conclusion: This system is suitable for clinical application.
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