This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Open AccessArticle
Flow Cytometric Immunophenotyping: Minimal Differences in Fresh and Cryopreserved Peripheral Blood Mononuclear Cells versus Whole Blood
by
Andrea Tompa
Andrea Tompa 1,2,
Junko Johansson
Junko Johansson 2,
Ulrika Islander
Ulrika Islander 2,3 and
Maria Faresjö
Maria Faresjö 2,*
1
Department of Clinical Diagnostics, School of Health and Welfare, Jönköping University, SE-551 11 Jönköping, Sweden
2
Division of Systems and Synthetic Biology, Department of Life Sciences, Chalmers University of Technology, SE-412 96 Gothenburg, Sweden
3
Department of Rheumatology and Inflammation Research, Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, SE-405 30 Gothenburg, Sweden
*
Author to whom correspondence should be addressed.
Biomedicines 2024, 12(10), 2319; https://doi.org/10.3390/biomedicines12102319 (registering DOI)
Submission received: 11 September 2024
/
Revised: 4 October 2024
/
Accepted: 6 October 2024
/
Published: 11 October 2024
Abstract
Background/Objectives: Flow cytometry is a convenient tool in immunophenotyping for monitoring the status of immunological conditions and diseases. The aim of this study was to investigate the effect of isolation and cryopreservation by flow cytometric analysis on subpopulations of CD4+ T helper (Th), T regulatory (Treg), CD8+ T cytotoxic (Tc), CD56+ NK, CD19+ B and monocytes. Freshly isolated and cryopreserved peripheral blood mononuclear cells (PBMCs) were compared to fresh whole blood. Methods: Peripheral blood was collected from healthy donors and prepared for flow cytometric analysis using the same panels of antibodies throughout the study. Results: Comparisons between fresh (F)- and cryopreserved (C)-PBMCs showed no major differences in percentages of CD4+, Th1, Th2 and CD4+CD25+CD127low Treg cells. No differences in percentage of CD8+ or subpopulations of naive/stem, central or effector memory cells were observed between F- and C-PBMCs. The percentage of CD56+ NK cells, CD19+ B cells or classical and nonclassical monocytes did not differ between F-and C-PBMCs either. On the contrary, whole blood had lower percentages of Th and NK cells but higher percentages of Th1, Th17, Th1Th17, Tregs, Tc and B cells compared to C-PBMCs, while it had a higher proportion of Tc compared to F-PBMCs. Conclusions: Flow cytometric immunophenotyping minimally differs between freshly isolated and cryopreserved PBMCs. This implies the possibility of cryostorage of cohorts for later analysis. Importantly, care must be taken when comparing results from whole blood with isolated and cryopreserved PBMCs. Collectively, these results can contribute to the standardization of flow cytometric protocols in both clinical and research settings.
Share and Cite
MDPI and ACS Style
Tompa, A.; Johansson, J.; Islander, U.; Faresjö, M.
Flow Cytometric Immunophenotyping: Minimal Differences in Fresh and Cryopreserved Peripheral Blood Mononuclear Cells versus Whole Blood. Biomedicines 2024, 12, 2319.
https://doi.org/10.3390/biomedicines12102319
AMA Style
Tompa A, Johansson J, Islander U, Faresjö M.
Flow Cytometric Immunophenotyping: Minimal Differences in Fresh and Cryopreserved Peripheral Blood Mononuclear Cells versus Whole Blood. Biomedicines. 2024; 12(10):2319.
https://doi.org/10.3390/biomedicines12102319
Chicago/Turabian Style
Tompa, Andrea, Junko Johansson, Ulrika Islander, and Maria Faresjö.
2024. "Flow Cytometric Immunophenotyping: Minimal Differences in Fresh and Cryopreserved Peripheral Blood Mononuclear Cells versus Whole Blood" Biomedicines 12, no. 10: 2319.
https://doi.org/10.3390/biomedicines12102319
Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details
here.
Article Metrics
Article metric data becomes available approximately 24 hours after publication online.