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A simple two‐step culture system for the large‐scale generation of mature and functional dendritic cells from umbilical cord blood CD34+ cells
Author(s) -
Balan Sreekumar,
Kale Vaijayanti P.,
Limaye Lalita S.
Publication year - 2009
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/j.1537-2995.2009.02231.x
Subject(s) - dendritic cell , cd34 , umbilical cord , cd14 , immunology , cord blood , immunotherapy , microbiology and biotechnology , biology , chemistry , stem cell , immune system
BACKGROUND: In vitro generated dendritic cells (DCs) are widely used as adjuvants in cancer immunotherapy. The major sources for DC generation are monocytes and CD34+ cells. CD34+‐derived DCs are less frequently used in clinical applications because it requires complex generation methods. Here a simple method for the large‐scale generation of mature functional DCs from umbilical cord blood–derived CD34+ cells is described. STUDY DESIGN AND METHODS: CD34+ cells were first expanded with a combination of early acting growth factors in a medium containing autologous plasma. In the second step the DC precursors were further either enriched by plastic adherence or sorted on a cell sorter and differentiated as DCs. DCs generated by both methods were compared for their morphology, phenotype, and different functional variables. RESULTS: This culture system provided a large‐scale expansion of CD34+ cells giving a mean fold increase of 615. The majority of the expanded cells were interstitial DC precursors, that is, CD14+‐positive cells. In vitro generated immature DCs could be matured into functional DCs by appropriate maturation stimuli. DCs generated by the plastic adherence method had a better cytokine profile and strong mixed leukocyte reaction compared to those generated by cell sorting. CONCLUSION: A two‐step culture system provides a large‐scale expansion of CD34+ cells with a preferential lineage commitment toward CD14+ cells. Enrichment of these precursors with a simple plastic adherence technique results in generation of large numbers of mature, functional DCs. This method of in vitro DC generation will have applications in cancer immunotherapy.