Primary monocyte-derived DCs were differentiated from CD14+ peripheral blood monocytes, and matured with LPS(100ng mL−1) for 2 days prior to use.10 (link) Primary human CD4+ T cells were positively isolated from CD14-depleted peripheral blood mononuclear cells (PBMCs) using CD4-conjugated magnetic beads and LS MACS cell separation columns (Miltenyi Biotech). Positively isolated CD4+ T cells were activated with 2% PHA (Invitrogen) for 2 days, washed and cultured in IL-2 (50 U/mL) containing RPMI supplemented with 10% FBS.
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Differentiation and Activation of Primary Immune Cells
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Other organizations : Boston University
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Variable analysis
independent variables
- Differentiation of primary monocyte-derived DCs from CD14+ peripheral blood monocytes
- Maturation of DCs with LPS (100ng mL-1) for 2 days
- Not explicitly mentioned
- Positive isolation of primary human CD4+ T cells from CD14-depleted peripheral blood mononuclear cells (PBMCs) using CD4-conjugated magnetic beads and LS MACS cell separation columns
- Activation of CD4+ T cells with 2% PHA for 2 days
- Culture of activated CD4+ T cells in IL-2 (50 U/mL) containing RPMI supplemented with 10% FBS
- Positive control: Not specified
- Negative control: Not specified
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