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Updated: May 23, 2026
Characterization of Human Monocyte Subsets by Whole Blood Flow Cytometry Analysis
Published on: October 17, 2018
Flow cytometric lymphocyte subset analysis using material from frozen whole blood
Iftikhar Alam1, David Goldeck, Anis Larbi
1Tübingen Aging and Tumour Immunology Group, Sektion für Transplantationsimmunologie und Immunohämatologie, University of Tübingen, Zentrum für MedizinischeForschung, Tübingen, Germany. iftikharalam@aup.edu.pk
Insights
Storing whole blood (WB) at -80°C is a viable alternative to cryopreserved peripheral blood mononuclear cells (PBMC) for immunological studies. This method preserves lymphocyte viability for T cell enumeration assays, even after extended storage.
Area of Science:
- Immunology
- Cellular Biology
- Biotechnology
Background:
- Multicenter immune monitoring often requires cryopreserved peripheral blood mononuclear cells (PBMC).
- PBMC isolation from whole blood before freezing is standard but requires specific laboratory facilities.
- Field conditions may lack the infrastructure for PBMC separation, posing logistical challenges.
Purpose of the Study:
- To assess the feasibility of using whole blood (WB) frozen at -80°C as a source of viable lymphocytes.
- To compare lymphocyte subsets from frozen WB with those from cryopreserved PBMC.
- To determine the suitability of frozen WB for immunological studies, particularly T cell enumeration.
Main Methods:
- Whole blood (WB) samples from five healthy donors were frozen at -80°C.
- Peripheral blood mononuclear cells (PBMC) were isolated and cryopreserved using standard methods.
- Flow cytometry was used to analyze CD4 and CD8 T lymphocyte percentages and subsets in both frozen WB and cryopreserved PBMC.
Main Results:
- Lymphocyte analysis from WB frozen up to 120 days showed results comparable to PBMC frozen up to 10 days.
- CD4 and CD8 T cell counts and subset percentages were similar between the two sample types within the tested durations.
- The viability and integrity of lymphocytes in frozen WB were maintained sufficiently for T cell enumeration.
Conclusions:
- Contrary to initial assumptions, whole blood (WB) frozen at -80°C can serve as an appropriate source of viable lymphocytes.
- This method offers a practical alternative for immunological and epidemiological studies where PBMC isolation facilities are unavailable.
- Frozen WB simplifies sample handling and storage for immune monitoring, expanding research possibilities in field settings.
Abstract:
Multicenter immune monitoring programs commonly rely on storing and shipping cryopreserved peripheral blood mononuclear cells (PBMC), isolated from whole blood before freezing. However, under many conditions in the field, facilities to separate PBMC are absent. Here, we investigate the feasibility of using whole blood (WB) frozen at -80°C as a source of viable lymphocytes for use in immunological studies. We compare the percentage of CD4 and CD8 T lymphocytes and their subsets from frozen WB with results from cryopreserved PBMC in five random healthy blood donors (three female, two male). We report that CD4 and CD8 values in lymphocytes from WB frozen up to 120 days were very similar to those of PBMC frozen up to 10 days. These data suggest that within the limits of parameters investigated in this study, contrary to our original assumptions, whole blood frozen at -80°C may in fact be an appropriate source of viable lymphocytes for T cell enumeration assays in immunological and epidemiological studies.
