Human dendritic cells were differentiated from human blood monocytes using the Human Monocyte-derived Dendritic Cell Differentiation kit (R&D Systems). Briefly, peripheral blood mononuclear cells were isolated from human blood (IRB approval # Project, No. 1711026297) followed by isolation of monocytes using Microbeads for human CD14+ isolation (Miltenyi Biotec), and incubation of 1 × 106 CD14+ cells/mL in differentiation media for 7 days at 37°C 5%CO2. Immature dendritic cells were incubated with 100 ng/mL LPS for 48 h and DAB2 expression assessed by Western blotting.
Human monocyte derived dendritic cell differentiation kit
Manufactured by R&D Systems
Sourced in United States
The Human Monocyte-derived Dendritic Cell Differentiation kit is a laboratory product that facilitates the differentiation of human monocytes into dendritic cells. It provides the necessary components and protocols to support this cellular differentiation process.
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2 protocols using human monocyte derived dendritic cell differentiation kit
1
Monocyte-derived Dendritic Cell Generation
2
Isolation and Differentiation of Human Dendritic Cells
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Blood samples were collected from Ahmed Maher teaching hospital blood bank, Egypt, in a vacutainer containing Heparin as anti-coagulant (Greiner bio-one, Australia) and processed immediately within 2 hours of collection.
The protocol was optimised for the preparation of PBMCs from human blood through using Ficoll-Hypaque density gradient centrifugation inaddition to using RPMI-10 complete medium (Gibco, United Kingdom) supplemented with penicillin/streptomycin (10 mg/ml), Glutamine (200 mM), 10% FCS, insulin and hydrocortisone.
In vitro culture and differentiation of human monocytes into dendritic cells DCs were achieved by using the Human Monocyte-derived Dendritic Cell Differentiation Kit (R&Dsystem, Minneapolis, MN, USA). This kit enables the generation of active mature DCs from human monocytes in 10 days (Amedei et al., 2011; Sabado and Bhardwaj, 2010).
The protocol was optimised for the preparation of PBMCs from human blood through using Ficoll-Hypaque density gradient centrifugation inaddition to using RPMI-10 complete medium (Gibco, United Kingdom) supplemented with penicillin/streptomycin (10 mg/ml), Glutamine (200 mM), 10% FCS, insulin and hydrocortisone.
In vitro culture and differentiation of human monocytes into dendritic cells DCs were achieved by using the Human Monocyte-derived Dendritic Cell Differentiation Kit (R&Dsystem, Minneapolis, MN, USA). This kit enables the generation of active mature DCs from human monocytes in 10 days (Amedei et al., 2011; Sabado and Bhardwaj, 2010).
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