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Phenotypic and genetic evaluation of human monocyte-derived dendritic cells generated from whole blood for immunotherapy

Authors
  • Hussein, Yousri M.1
  • Hendawy, Doaa M.1
  • Alghamdy, Abdalrahman N.2
  • Raafat, Nermin1
  • 1 Zagazig University, Zagazig, Egypt , Zagazig (Egypt)
  • 2 Taif University, Taif, Saudi Arabia , Taif (Saudi Arabia)
Type
Published Article
Journal
Egyptian Journal of Medical Human Genetics
Publisher
Springer Berlin Heidelberg
Publication Date
May 25, 2021
Volume
22
Issue
1
Identifiers
DOI: 10.1186/s43042-021-00168-7
Source
Springer Nature
Keywords
License
Green

Abstract

BackgroundDendritic cells (DCs) recognize different pathogens and cancer cells and activate the adaptive immune response. The generation of effective DC-based cancer vaccines depends on the appropriate differentiation of monocytes in vitro. This study aimed to standardize a protocol for the in vitro differentiation of human peripheral blood monocytes into immature DCs upon treatment with growth factors and generate monocyte-derived DCs (MoDCs). Peripheral blood mononuclear cells were separated from peripheral blood. After monocyte enrichment by plastic adhesion, monocytes were cultured for 6 days in the presence of granulocyte-macrophage colony-stimulating factor and interleukin-4 to generate immature DCs. The cells were examined by microscopy. Using flow cytometry, DCs were evaluated for the expression of the CD83 and HLA-DR surface antigens, for the uptake of fluorescein isothiocyanate conjugated dextran, and also for the expression of CD80 and CD86 mRNA.ResultsCD80 and CD86 genes expression was upregulated at day six and exhibited a significant difference (P < 0.05). DCs showed positive expression of the CD83 and HLA-DR surface antigens by flow cytometry and FITC-conjugated dextran uptake.ConclusionThis study represents a preliminary trial to generate immature MoDCs in vitro from blood monocytes collected by the flask adherence method. It offers a panel of surface markers for DCs characterization and provides Immature DCs for experimental procedures after 6 incubation days.

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