DCs differentiation and stimulation

PBMC were isolated from buffy coats by density gradient centrifugation using Biocoll (BIOCHROM). Monocytes were isolated from PBMC using MACS anti-CD14 microbeads and a Midi-MACS® magnetic cell sorting device (both from Miltenyi Biotec, Bergisch-Gladbach,Germany). Cells were cultured in RPMI 1640 medium (GIBCO BRL) supplemented with 2 mM L-glutamine (Sigma), 1% (vol/vol) non-essential amino acids, 100 mM sodium pyruvate, 50 U/ml of penicillin and 50 mg/ml of streptomycin (Gibco BRL) containing 10% (vol/vol) FCS (Hyclone). Differentiation of monocytes into dendritic cells was promoted by addition of granulocyte-macrophage colony-stimulating factor (GMCSF 1000U/ml, Chemicon) and recombinant IL-4 (1000U/ml, R&D Systems) for 5 days. DCs activation was induced by lipopolisaccharide (LPS 1microgram/ml, Sigma, St. Louis, MO), by Curdlan (100 microgram/ml, Wako), by R848 and yeast RNA and by yeast cells in different conditions of culture.

• biomaterial type
• lipopolysaccharide,
• streptomycin,
• L-Glutamine,
• ribonucleic acids,
• amino acids,
• resiquimod,
• monocyte,
• buffy coat,
• Peripheral Blood Mononuclear Cell,
• Interleukin-4,
• curdlan,
• Saccharomyces cerevisiae

created over 16 years ago (2 March 2009)    last modified over 13 years ago (28 September 2011)   [ RDF ]   [ RelFinder ]