Transcription profiling of human immature dendritic cells treated with Lipopolysaccharide (LPS) vs Taxol (paclitaxel)

ABSTRACT: Generation of monocyte-derived dendritic cells (DC). Immature DC were prepared from peripheral blood mononuclear cells (PBMCs) isolated from whole blood of healthy donors. A minimum of 3 individual donor blood was used for each experiment. PBMCs were incubated with anti-CD14 microbeads (Miltenyi Biotec, Bergish Gladbach, Germany) as specified in the manufacturer's instructions. The isolated monocytes were resuspended at 10^6 cells/ml in medium supplemented with 10% human AB serum, 100 ng/ml GM-CSF (Leucomax; Novartis, Camberley, UK) and 50 ng/ml IL-4 (Peprotech EC Ltd, London, UK), cultured in tissue culture flasks. On day 7 the cells were defined as immature DC with low/intermediate HLA-DR expression. Microarray analysis of the effects of Paclitaxel and LPS on DC. Total RNA was extracted from day 7 DC which had been treated for 2 h with 1 ug/ml LPS or 100 uM paclitaxel for 2 h then washed and returned to culture for 24 h. This was used as a template to generate Cy3-labelled cRNA, using the Low RNA Input Linear Amplification Kit (Agilent). This was used as a probe on the Whole Human Genome Microarray (4x44K) slide (Agilent). Slides were scanned using the Agilent scanner and data extracted using Feature Extraction Software 9.5.3 (Agilent). Subsequent data analysis was performed using GeneSpring GX software.

ORGANISM(S): Homo sapiens

SUBMITTER: Richard Morgan 

PROVIDER: E-MEXP-2465 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress