Gene Expression Profiles from Peripheral Blood Mononuclear Cells Are Sensitive to Short Processing Delays.
Author(s): Barnes MG, Grom AA, Griffin TA, Colbert RA, Thompson SD
Publication: Biopreserv Biobank, 2010, Vol. 8, Page 153-162
PubMed ID: 21743826 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to determine the effects of delayed processing on transcriptome analysis of PBMC using Affymetrix HG U133 Plus 2.0 microarrays. After isolation, PBMC were stored at -80 degrees C until RNA extraction. Each time-point in the initial analysis included only 3 specimens, but the confirmation study included 276 specimens.
Summary of Findings:
When blood was stored at room temperature for 4 h prior to harvesting PBMC, 307 transcripts displayed a >2 fold increase, and 46 transcripts displayed a >2 fold decrease in levels compared to levels in specimens harvested immediately after phlebotomy. Storage of whole blood at room temperature for 2 h prior to harvesting also led to significant changes in transcript levels. When the lists of transcripts affected by a 4 h delay in processing were compared to those in a previous study of the effect of an overnight room temperature delay before PBMC harvesting, only 17% of the transcripts were common to the two studies. However, the 353 transcripts that were altered after a 4 h room temperature delay prior to harvesting PBMC (3 specimens) showed similar trends after 60-315 min delays in freezing of PBMC from 276 additional specimens, including those from juvenile idiopathic arthritic and normal individuals, and alterations were independent of diagnosis.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Normal
- Arthritis
Platform:
Analyte Technology Platform RNA DNA microarray Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Storage duration 0 h
2 h
4 h
1-5.25 h
Overnight
Preaquisition Diagnosis/ patient condition Juvenile idiopathic arthritis
Normal