Combined therapy composition and combined therapy method for treating cancers
一种免疫治疗、靶向治疗剂的技术,应用在化学仪器和方法、药物组合、基因治疗等方向
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[0468] Enrichment of Human Dendritic Cells (DC) from PBMC
[0469] Human PBMCs were prepared from erythrocytes obtained from healthy voluntary donors by Ficoll centrifugation. Dendritic cells were enriched from magnetic beads (Miltenyi Biotec Inc. San Diego, CA) with a mixture of anti-CD3, anti-CD19, anti-CD20, anti-CD14 and anti-CD16 antibodies from human PBMCs by using negative subtraction. Enriched DCs were stained with goat anti-mouse FITC (lineage), HLA-DR-APCCy7, CD123-BV421 and CD11C-APC. Stained cells were analyzed on BDLSR Fortessa (BD Biosciences). Anti-CD3 mAb, anti-CD4 mAb, anti-CD11C mAb, anti-CD19 mAb, anti-CD14 mAb, anti-CD16 mAb, anti-CD123 mAb were purchased from BD Biosciences, CA or Biolegend, San Diego , CA.
[0470] Stimulation of enriched human DCs and expression of cytokines
[0471] Will be 1-2 x 10 5 Enriched DCs were seeded in 96-well plates in 100 μl of medium, 100 μl of diluted stimulators including BDs100 (resiquimod) were added to the pla...
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