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| Status |
Public on Jun 15, 2009 |
| Title |
Gene expression profiles in human keratinocytes exposure to ultrafine, fine, and submicron TiO2 particles |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Gene expression profiling of the human keratinocytes cell line (HaCaT) exposure to ultrafine, fine, and submicron TiO2 particles were employed to gain insights into the molecular events.
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| Overall design |
Three types of bulk TiO2 anatase particles, ST-01(average primary particle size= 7nm, specific surface area = 316m2/g), ST-21(20nm and 66m2/g), and ST-41(200nm and 10m2/g) used in this study were purchased from Ishihara Sangyo Kaisha Ltd., Japan. TiO2 particles were dispersed and size-fractionated in Dulbecco’s modified Eagle’s medium (DMEM) . TiO2 particles was dispersed in heat-inactivated fetal bovine serum (FBS) and centrifuged at 16,000 x g for 20 min. Removing the supernatant, the pellet was washed with DMEM, and centrifuged at 16,000 x g for 20 min. The supernatant was removed and dispersed in fresh DMEM with 10 % FBS medium and antibiotic/antifungal-agent (100 units/ml of penicillin, 100 μg/ml of streptomycin, and 250 ng/ml of amphotericin B) defined as DMEM-FBS medium, and centrifuged at 8,000 x g for 20 min. Removing the supernatant, the pellet was vortexed with fresh DMEM-FBS medium and centrifuged at 4,000 x g for 20min. The supernatant of 4,000 x g fraction was collected. Continuously, the centrifugal force was reduced to 2,000 x g, 1,000 x g and 500 x g. Each supernatant of 2,000 x g, 1,000 x g, and 500 x g fraction was collected. The 500 x g fraction supernatant was 0.8 µm membrane-filtered. The concentrations and the secondary average diameter of TiO2 particles in the supernatant fractions were individually characterized using X-ray diffraction (XRD), dynamic light scattering (DLS), and transmission electron microscopy (TEM) at 120 kV. The samples were defined as T-7 (ST-01, 4,000 x g fraction), T-20 (ST-21, 1,000 x g fraction), and T-200 (ST-41, 500 x g fraction), and were utilized for the following DNA microarray analysis. Human keratinocyte cells (HaCaT) purchased from the German Cancer Research Center (DKFZ) were cultured in DMEM-FBS medium at 37 °C in a humidified atmosphere of 5% CO2 and 95% air The passage numbers 2–10 were used in the experiments, and cells were seeded at a density of approximately 2 x 10^5 cells/ml. All experiments were performed in triplicate (n=3).
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| Contributor(s) |
Fujita K, Iwahashi H |
| Citation(s) |
19695317 |
| Submission date |
Jun 04, 2009 |
| Last update date |
Jan 23, 2019 |
| Contact name |
Katsuhide Fujita |
| E-mail(s) |
ka-fujita@aist.go.jp
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| Organization name |
National Institute of Advanced Industrial Science and Technology
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| Department |
Research Institute of Science for Safety and Sustainability
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| Street address |
Onogawa 16-1
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| City |
Tsukuba |
| ZIP/Postal code |
3058569 |
| Country |
Japan |
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| Platforms (1) |
| GPL6480 |
Agilent-014850 Whole Human Genome Microarray 4x44K G4112F (Probe Name version) |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA116371 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE16425_RAW.tar |
324.3 Mb |
(http)(custom) |
TAR (of TXT) |
| Processed data included within Sample table |
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