Related Experiment Video
Updated: May 9, 2026
Cryopreservation and Bioenergetic Evaluation of Human Peripheral Blood Mononuclear Cells
Published on: October 20, 2023
Temperature fluctuations during deep temperature cryopreservation reduce PBMC recovery, viability and T-cell function
Anja Germann1, Young-Joo Oh, Tomm Schmidt
1(a)Fraunhofer Institute for Biomedical Engineering, Ensheimerstr. 48, 66386 St. Ingbert, Germany.
Insights
Minimizing temperature fluctuations during cryopreservation of peripheral blood mononuclear cells (PBMCs) is crucial. A protective hood system preserves cell viability, recovery, and T-cell functionality, vital for immune-based therapy trials.
Area of Science:
- Immunology
- Cryobiology
- Biobanking
Background:
- Cryopreserved peripheral blood mononuclear cells (PBMCs) are essential for evaluating immune-based therapies.
- Standardized cryopreservation is needed for reproducible results in multicenter trials.
- Suboptimal storage conditions can compromise cell viability and functionality.
Purpose of the Study:
- To develop and evaluate a storage approach minimizing temperature fluctuations.
- To assess the impact of temperature fluctuations on PBMC viability, recovery, and T-cell function.
- To compare a protective hood system against standard handling during cryopreservation.
Main Methods:
- Comparison of PBMC storage with and without temperature fluctuations.
- Utilizing a protective hood system to mitigate temperature rises during sample handling.
- Assessing cell viability, recovery rates, and antigen-specific T-cell responses.
Main Results:
- Cyclical temperature shifts significantly reduce PBMC viability, recovery, and immune response.
- The protective hood system maintained cell viability and recovery comparable to stable storage.
- T-cell functionality was considerably increased when using the protective hood system.
Conclusions:
- Temperature fluctuations during cryopreservation negatively impact PBMC integrity and function.
- A protective hood system effectively preserves PBMC quality for immunological assays.
- Optimal sample storage conditions are critical for reliable clinical vaccine trial outcomes.
Abstract:
The ability to analyze cryopreserved peripheral blood mononuclear cell (PBMC) from biobanks for antigen-specific immunity is necessary to evaluate response to immune-based therapies. To ensure comparable assay results, collaborative research in multicenter trials needs reliable and reproducible cryopreservation that maintains cell viability and functionality. A standardized cryopreservation procedure is comprised of not only sample collection, preparation and freezing but also low temperature storage in liquid nitrogen without any temperature fluctuations, to avoid cell damage. Therefore, we have developed a storage approach to minimize suboptimal storage conditions in order to maximize cell viability, recovery and T-cell functionality. We compared the influence of repeated temperature fluctuations on cell health from sample storage, sample sorting and removal in comparison to sample storage without temperature rises. We found that cyclical temperature shifts during low temperature storage reduce cell viability, recovery and immune response against specific-antigens. We showed that samples handled under a protective hood system, to avoid or minimize such repeated temperature rises, have comparable cell viability and cell recovery rates to samples stored without any temperature fluctuations. Also T-cell functionality could be considerably increased with the use of the protective hood system compared to sample handling without such a protection system. This data suggests that the impact of temperature fluctuation on cell integrity should be carefully considered in future clinical vaccine trials and consideration should be given to optimal sample storage conditions.
