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

ABSTRACT: 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.

SUBMITTER: Andriamboavonjy L

PROVIDER: S-EPMC10754832 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature

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

Andriamboavonjy Lovatiana L MacDonald Adam A Hamilton Laura K LK Labrecque Marjorie M Boivin Marie-Noёlle MN Karamchandani Jason J Stratton Jo Anne JA Tetreault Martine M

Scientific reports 20231228 1

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 ...[more]

PMID: 38155174

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

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

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