Related Experiment Video
Updated: May 31, 2026
09:08
Isolating Human Peripheral Blood Mononuclear Cells and CD4+ T cells from Sézary Syndrome Patients for Transcriptomic Profiling
Published on: October 14, 2021
Gene Expression Profiles from Peripheral Blood Mononuclear Cells Are Sensitive to Short Processing Delays
Michael G Barnes1, Alexei A Grom, Thomas A Griffin
1Division of Rheumatology, Cincinnati Children's Hospital Medical Center and University of Cincinnati College of Medicine, Cincinnati, Ohio.
Biopreservation and Biobanking
|July 12, 2011
Summary
Delayed processing of peripheral blood mononuclear cells (PBMCs) significantly alters gene expression. Ensure timely sample handling in translational studies to accurately reflect in vivo biology.
Area of Science:
- Molecular Biology
- Immunology
- Genomics
Background:
- Accurate gene expression analysis in peripheral blood requires samples processed quickly to avoid ex vivo artifacts.
- Peripheral blood mononuclear cells (PBMCs) are crucial for understanding immune responses and disease states.
Purpose of the Study:
- To investigate the impact of processing delays on global gene expression patterns in human PBMCs.
- To identify specific genes and pathways affected by ex vivo sample handling time.
Main Methods:
- PBMC RNA was isolated and stabilized immediately or after a 4-hour delay from 3 individuals.
- Gene expression profiling was performed using Affymetrix HG U133 Plus 2.0 GeneChips.
- Differential gene expression analysis identified significant changes (≥2-fold, P < 0.05) after delay.
Main Results:
- A 4-hour delay in PBMC processing resulted in 307 probe sets with increased expression and 46 with decreased expression.
- Affected genes are involved in critical inflammatory, immunologic, and cancer pathways.
- Expression trends were confirmed in an independent dataset of 276 PBMC arrays with variable processing times.
Conclusions:
- Processing delays significantly alter PBMC gene expression profiles, impacting translational research.
- Time from sample acquisition to RNA stabilization is a critical factor for reliable gene expression analysis.
- Standardized, timely processing protocols are essential for reproducible PBMC gene expression studies.
