A large number of mature and functional dendritic cells can be efficiently generated from umbilical cord blood-derived mononuclear cells by a simple two-step culture method.
BACKGROUND
Advances in the past two decades in dendritic cell (DC) biology paved the way to exploit them as a promising tool in cancer immunotherapy. The prerequisite for DC vaccine preparations is large-scale in vitro generations of homogeneous, mature, and functional DCs. Frequent improvements are being made in the existing in vitro DC production protocols to achieve this goal. In our previous study we reported a large-scale generation of mature, functional DCs from umbilical cord blood (UCB) CD34+ cells. Here we report that this method can be used for the efficient generation of DCs from UCB mononuclear cells (MNCs) and thus the hematopoietic stem cell isolation step is not essential.


METHODS
MNCs or CD34+ cells isolated from the same cord blood (CB) samples were used for the generation of DCs. DCs were characterized for morphology, phenotype, and functional assays including antigen uptake, chemotaxis, and mixed leukocyte reaction. Similarly DCs generated from the MNCs of same fresh and frozen CB units were compared.


RESULTS
The morphologic, phenotypic, and functional characterization of the DCs generated from various sets show that they were comparable in nature irrespective of the starting population used.


CONCLUSIONS
We conclude that the CD34+ isolation step is not essential for the generation of mature, functional DCs and thus can be eliminated. More importantly, we show that DCs can be generated with equal efficiency from the MNCs of frozen CB units. Our culture method will be useful for exploiting the potential of UCB as an additional source for allogeneic DCs in the clinical settings.
Balan
Sreekumar
S
National Centre for Cell Science, Pune University Campus, Pune, India.
Kale
Vaijayanti P
VP
Limaye
Lalita S
LS
D000941:Antigens / Q000378:metabolism; D018952:Antigens, CD34 / Q000378:metabolism; D001793:Blood Preservation; D019496:Cancer Vaccines; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000276:immunology; D002634:Chemotaxis, Leukocyte / Q000276:immunology; D015925:Cryopreservation; D003713:Dendritic Cells / Q000166:cytology*; D005312:Fetal Blood / Q000166:cytology*; D006801:Humans; D016130:Immunophenotyping; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000378:metabolism; D007959:Lymphocyte Culture Test, Mixed; D054400:Receptors, CCR7 / Q000378:metabolism
D000941:Antigens; D018952:Antigens, CD34; C515370:CCR7 protein, human; D019496:Cancer Vaccines; D054400:Receptors, CCR7