Author:
Andriamboavonjy Lovatiana,MacDonald Adam,Hamilton Laura K.,Labrecque Marjorie,Boivin Marie-Noёlle,Karamchandani Jason,Stratton Jo Anne,Tetreault Martine
Abstract
AbstractPreserving 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.
Funder
Schlumberger Foundation
Faculty of Medicine, University of Montreal
FRQS/Parkinson's Quebec
The Michael J. Fox Foundation for Parkinson’s Research
Aligning Science Across Parkinson’s (ASAP) initiative
Fond de recherche du Québec - Santé
Fondation Courtois
American Parkinson’s Disease Association
Publisher
Springer Science and Business Media LLC