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A global look into human T cell subsets before and after cryopreservation using multiparametric flow cytometry and
Jennifer Lemieux1, Christine Jobin1, Carl Simard2
1Recherche et Développement, Héma-Québec, Québec, QC G1V-5C3, Canada; Biochimie, Microbiologie et Bio-informatique, Faculté des Sciences et de Génie, Université Laval, Québec, QC G1V-0A6, Canada.
Journal of Immunological Methods
|May 1, 2016
Summary
Cryopreservation impacts human lymphocytes, crucial for cellular therapies. A 1-hour rest period post-thaw optimizes T cell subset detection, enhancing quality control for cancer treatments.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Cryopreservation of human lymphocytes is vital for cellular therapies.
- T cells show promise in cancer therapy due to their cytotoxic and regulatory functions.
- Monitoring cryopreservation effects on lymphocyte distribution is key for quality control.
Purpose of the Study:
- To compare phenotypes of fresh human mononuclear cells with those post-cryopreservation.
- To evaluate the impact of different rest periods (0h, 1h, 24h) after thawing on T cell subsets.
- To assess the utility of SPADE and viSNE software for monitoring cryopreservation effects.
Main Methods:
- Flow cytometry with eight-color antibody panels.
- Analysis of naive, memory effector, regulatory, and helper T cell subsets.
- Comparison of data using conventional gating, SPADE, and viSNE clustering software.
Main Results:
- T cells are more sensitive to cryopreservation than other cell types.
- A 1-hour rest period post-thaw improved the detection of certain cell markers compared to fresh samples.
- SPADE and viSNE effectively monitored PBMC and T cell subset distribution changes after cryopreservation.
Conclusions:
- SPADE and viSNE are efficient tools for monitoring cryopreservation impacts on PBMC and T cell subsets.
- A 1-hour rest period post-thaw is beneficial for accurate cell marker detection.
- Cryopreservation significantly affects lymphocyte distribution, necessitating optimized protocols for cellular therapy quality control.
