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X-ray exposure of peripheral blood mononuclear cells

ABSTRACT: Peripheral blood mononuclear cells (PBMCs) were isolated using Histpaque-1077 (Sigma-Aldrich, Belgium) from 10 healthy donors (5 males, 5 females, aged 23-50). Isolated cells were resuspended at a density of 10^6 cells/ml and allowed to equilibrate to culture conditions at 37°C in a humidified 5% CO2 atmosphere. PBMCs were then exposed to 0.0 (sham), 0.1 or 1.0 Gy of X-rays (250 keV - 1 mA, 1 mm Cu) at a dose rate of 0.26 Gy/min. After irradiation, cells were incubated at 37°C in a humidified 5% CO2 atmosphere for 8 h before RNA extraction using the AllPrep DNA/RNA/protein Mini kit (Qiagen). Two weeks later, the experiment was repeated using fresh PBMCs from the same donors, resulting in a total of 60 RNA samples. Total RNA (250 ng) was used for hybridisation to GeneChip Human Gene 1.0 ST arrays (Affymetrix) using the WT expression kit (Ambion), GeneChip Terminal labeling kit (Affymetrix) and the GeneChip Hybridization, Wash and Stain kit (Affymetrix) according to the manufacturer's recommendations.

ORGANISM(S): Homo sapiens

SUBMITTER: Roel Quintens

PROVIDER: E-MTAB-3463 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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