Related Experiment Videos
A clinically applicable method for the ex vivo generation of antigen-presenting cells from CD34+ progenitors
K M Ardeshna1, C P Corney, S J Ings
1Department of Haematology, Royal Free and University College Medical School, London, UK. k.ardeshna@ucl.ac.uk
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-alpha and rhIL-4. Large-scale cultures were performed in Stericell 12 inch x 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.