Comparative analysis of methods to reduce activation signature gene expression in PBMCs

Preserving the in vivo cell transcriptome is essential for accurate profiling, yet factors during cell isolation including time ex vivo and temperature induce artifactual gene expression, particularly in stress-responsive immune cells. In this study, we investigated two methods to mitigate ex vivo a...

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Bibliographic Details
Published in:Scientific reports 2023-12, Vol.13 (1), p.23086-23086, Article 23086
Main Authors: Andriamboavonjy, Lovatiana, MacDonald, Adam, Hamilton, Laura K., Labrecque, Marjorie, Boivin, Marie-Noёlle, Karamchandani, Jason, Stratton, Jo Anne, Tetreault, Martine
Format: Article
Language:English
Subjects:
631/250
631/80
Blood
Chemokines
Cold Temperature
Comparative analysis
Gene expression
Gene Expression Profiling - methods
Humanities and Social Sciences
Leukocytes (mononuclear)
Leukocytes, Mononuclear - metabolism
multidisciplinary
Peripheral blood mononuclear cells
Science
Science (multidisciplinary)
Temperature
Transcriptome
Transcriptomes
Transcriptomics
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Summary:Preserving the in vivo cell transcriptome is essential for accurate profiling, yet factors during cell isolation including time ex vivo and temperature induce artifactual gene expression, particularly in stress-responsive immune cells. In this study, we investigated two methods to mitigate ex vivo activation signature gene (ASG) expression in peripheral blood mononuclear cells (PBMCs): transcription and translation inhibitors (TTis) and cold temperatures during isolation. Comparative analysis of PBMCs isolated with TTis revealed reduced ASG expression. However, TTi treatment impaired responsiveness to LPS stimulation in subsequent in vitro experiments. In contrast, cold isolation methods also prevented ASG expression; up to a point where the addition of TTis during cold isolation offered minimal additional advantage. These findings highlight the importance of considering the advantages and drawbacks of different isolation methods to ensure accurate interpretation of PBMC transcriptomic profiles.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-023-49611-2