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Series GSE22022 Query DataSets for GSE22022
Status Public on Aug 31, 2010
Title Microarray profiling of expression patterns of fibroblasts from different skin types
Organism Homo sapiens
Experiment type Expression profiling by array
Summary Adult human fibroblasts derived from the dermis of 4 mm punch biopsies taken from the lower backs of 15 healthy subjects of 3 different phototypes (types I, III and VI). This study was approved by the Human Research Ethics Committee of Hamburg. Subjects with type I skin were #4, 12, 15, 19 and 26; subjects with type III skin were # 6, 9, 13, 14 and 23, and subjects with type VI skin were # 7, 8, 16, 17 and 28. The fibroblasts were cultured from the biopsies and were grown in monolayer culture in high glucose Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% L-glutamine plus 1% penicillin and streptomycin at 37°C in a humidified 5% CO2 atmosphere. Fibroblasts were subcultured using routine methods and were used at passages 3 to 6.
 
Overall design Oligo-cDNA microarray hybridization was performed according to the National Cancer Institute in-house protocol, as detailed previously (Yamaguchi et al., 2008). Briefly, total RNAs were prepared from cultured fibroblasts using an RNeasy mini kit (Qiagen, Valencia, CA). The quality (purity and integrity) and quantity of each total RNA preparation was measured using a Nanodrop ND-1000 spectrophotometer (Thermo Scientific, Wilmington, DE). A universal human reference RNA (Stratagene, La Jolla, CA) was used as a control. cDNA samples generated from RNA samples and purified before the coupling reaction were labeled with Cy3 (for fibroblast samples) or Cy5 (for reference RNA) mono-reactive dyes (GE Healthcare, Piscataway, NJ), and were hybridized simultaneously on an oligo-DNA chip (Hs-OperonV3.0-v1p24, p27, p31) overnight at 42°C. Two fluorescent intensities of the oligo-DNA chip were scanned using a microarray scanner (GenePix 4000B; Axon Instruments Inc., Molecular Devices Corp., Sunnyvale, CA). Differential gene expression was profiled using Genepix Pro 5.0 software and was analyzed by Miltenyi Biotec (Bergisch Gladbach, Germany).
 
Contributor(s) Hearing VJ, Choi W
Citation(s) 20736300
Submission date May 27, 2010
Last update date Mar 22, 2012
Contact name Vincent Hearing
E-mail(s) hearingv@nih.gov
Organization name National Institutes of Health
Department National Cancer Institute
Lab Laboratory of Cell Biology
Street address Bldg 37 Rm 2132
City Bethesda
State/province MD
ZIP/Postal code 20892
Country USA
 
Platforms (1)
GPL3779 NCI/ATC Hs-OperonV3
Samples (15)
GSM547580 Skin Type I, Patient 12
GSM547581 Skin Type I, Patient 15
GSM547582 Skin Type I, Patient 19
Relations
BioProject PRJNA127443

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE22022_RAW.tar 50.0 Mb (http)(custom) TAR (of GPR)
Raw data provided as supplementary file
Processed data included within Sample table

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