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Updated: May 29, 2025
Author Spotlight: Preservation of Bioenergetic Parameters in Peripheral Blood Mononuclear Cells After Cryopreservation
Published on: October 20, 2023
Impact of cryopreservation on immune cell metabolism as measured by SCENITH
Curtis Luscombe1, Eben Jones1, Michaela Gregorova1
1School of Cellular and Molecular Medicine, University of Bristol, Bristol, BS8 1TD, United Kingdom.
Insights
Cryopreservation impacts immune cell metabolism, impairing mitochondrial function and favoring glycolysis. However, the SCENITH technique can still profile immune cell bioenergetics in cryopreserved samples.
Area of Science:
- Immunology
- Cellular Metabolism
- Flow Cytometry
Background:
- Immune cell function is intrinsically linked to cellular metabolism.
- Dysfunctional immune responses are increasingly studied through immunometabolic correlates.
- Cryopreservation is common for clinical samples but may induce metabolic alterations.
Purpose of the Study:
- To investigate the effects of cryopreservation on immune cell metabolism.
- To assess the utility of SCENITH for metabolic profiling of cryopreserved immune cells.
- To understand how cryopreservation influences immune cell bioenergetics.
Main Methods:
- Utilized SCENITH, a flow cytometry-based method for ex vivo metabolic profiling.
- Analyzed immune cell subsets from cryopreserved clinical samples.
- Compared metabolic profiles of fresh versus cryopreserved immune cells.
Main Results:
- Cryopreservation induced a time-dependent metabolic artefact, favoring glycolysis and impairing oxidative phosphorylation.
- Activated T cells showed limited metabolic reprogramming capacity post-cryopreservation.
- SCENITH successfully differentiated bioenergetic profiles of immune cell populations despite cryopreservation artefacts.
Conclusions:
- Cryopreservation can lead to mitochondrial dysfunction and metabolic artefacts in immune cells.
- SCENITH remains valuable for studying immune cell metabolism in cryopreserved samples.
- Careful sample processing and storage are crucial to mitigate cryopreservation's impact on immunometabolic analyses.
Abstract:
The dynamic functioning of immune cells is regulated by cellular metabolic processes, and there is growing interest in the study of immunometabolic correlates of dysfunctional immune responses. SCENITH is a novel flow cytometry-based technique that allows for ex vivo metabolic profiling of immune cells within heterogeneous samples. Cryopreservation of clinical samples is frequently undertaken to facilitate high throughput processing and longitudinal analyses of immune responses, but is thought to lead to cellular metabolic dysfunction. We aimed to investigate the impact of cryopreservation on immune cell metabolism, harnessing SCENITH's unique ability to describe the divergent bioenergetic characteristics of distinct immune cell subsets. We demonstrate that upon activation, T cells are unable to sufficiently/readily undergo metabolic reprogramming. Additionally, we find that cryopreservation introduces a time-dependent metabolic artefact that favours glycolysis and impairs oxidative phosphorylation, suggesting that cryopreservation results in mitochondrial dysfunction. Despite this artefact, SCENITH was still able to reveal the distinct bioenergetic profiles of contrasting immune cells populations following cryopreservation. Whilst SCENITH can provide valuable information about immune cell metabolism even in cryopreserved samples, our findings have important implications for the design of future studies. Investigators should carefully consider how to process and store clinical samples to ensure that cryopreservation does not confound analyses, particularly where longitudinal sampling is required.
