A clinically applicable method for the ex vivo generation of antigen-presenting cells from CD34+ progenitors.
OBJECTIVE
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.


CONCLUSIONS
This system is suitable for clinical application.
Ardeshna
K M
KM
Department of Haematology, Royal Free and University College Medical School, London, UK. k.ardeshna@ucl.ac.uk
Corney
C P
CP
Ings
S J
SJ
Watts
M J
MJ
Linch
D C
DC
Devereux
S
S
D000818:Animals; D017951:Antigen Presentation; D018952:Antigens, CD34; D002417:Cattle; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology*; D005434:Flow Cytometry; D019650:Hematopoietic Stem Cell Mobilization / Q000379:methods; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000276:immunology*; D006801:Humans
D018952:Antigens, CD34