Effects of blood sample storage time, temperature, anti-coagulants and blood stabiliser on lymphocyte phenotyping.
Author(s): Lao Y, Quach A, Perveen K, Hii C, Ferrante A
Publication: Pathology, 2024, Vol. 56, Page 571-576
PubMed ID: 38403560 PubMed Review Paper? No
Purpose of Paper
This paper compared the stability of lymphocyte subpopulations among lithium heparin and EDTA blood and blood preserved with Streck Cell Preservative during storage at 4°C and room temperature for up to 48 h.
Conclusion of Paper
Storage of blood at room temperature for 24 or 48 h led to significant changes in the percentages of CD8+, CD3+ and CD16+/CD56+ NK cell populations in lithium heparin blood and changes in the percentages of CD2+, CD3+, CD4+, CD8+, CD16+/CD56+ natural killer (NK), CD19+ and CD20+ cells in EDTA blood. However, while storage at 4°C rather than room temperature led to changes in more lymphocyte subpopulations in lithium heparin blood (CD8+, CD4+, CD19+, CD20+, HLA-DR+ CD3+ and CD2+ cells), fewer lymphocyte subpopulations were affected in EDTA blood (CD8+ and CD4+ cells, only). When Streck Cell Preservative was added to EDTA blood, there was a higher percentage of CD19+ and a lower percentage of CD4+/CD3+ cells after 4 h at room temperature but not 24 or 48 h, and a change in the percentage of CD2+ and CD20+ cells after 24 or 48 h storage at room temperature. None of the lymphocyte subpopulations were significantly affected by storage of blood with Streck Cell Preservative at 4°C for up to 48 h.
The authors conclude that storage of either lithium heparin or EDTA blood for ≥24 h has a significant impact on lymphocyte subpopulations; while more lymphocyte subtypes are affected by storage of lithium heparin blood at 4°C than room temperature, the lymphocyte population is more stable in EDTA blood at 4°C than room temperature. Importantly, the addition of Streck Cell Preservative to blood partially stabilized lymphocyte subpopulations.
Studies
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Study Purpose
This study compared the stability of lymphocyte subpopulations among lithium heparin and EDTA blood and blood preserved with Streck Cell Preservative during storage at 4°C and room temperature for up to 48 h. Venous blood was collected from 15 healthy volunteers into lithium heparin or EDTA Vacutainer tubes. Within 2 h of venipuncture, 1 mL of blood from EDTA tubes was preserved with Streck Cell Preservative. The blood in lithium heparin (15 specimens) and EDTA tubes (12 specimens) and the blood preserved with Streck Cell Preservative (14 specimens) were stored at 4°C and room temperature for 4 (EDTA and Streck Cell Preservative only), 24 and 48 h before flow cytometry. Lymphocytes were phenotyped by flow cytometry using antibodies against CD2, CD3, CD4, CD8, CD19, CD20, CD16, CD56 and HLA-DR.
Summary of Findings:
After 24 h of storage at room temperature, lithium heparin blood had a higher mean percentage of CD8+ cells (p=0.017), and after 48 h there was a higher mean percentage of CD8+ (P<0.001) and CD3+ cells (p<0.001) and a decrease in the percentage CD16+/CD56+ NK cells (p<0.001). After 24 h of storage at 4°C, lithium heparin blood had a higher mean percentage of CD8+ (p=0.015) and CD4+ cells (p=0.010), and after 48 h at 4°C there was a higher mean percentage of CD8+ (P<0.001), CD4+ (p<0.001), CD19+ (p=0.003), CD20+ (p<0.001) and HLA-DR+ CD3+(p=0.004) cells and a decrease in CD2+ cells (p<0.001).
Blood stored in EDTA tubes at room temperature for 24 h or 48 h had a higher mean percentage of CD2+ (p<0.001, both), CD3+ (p=0.013 and p<0.001, respectively), CD4+ (not significant, NS and p=0.002, respectively) and CD8+ cells (p<0.001, both) and decreased percentages of CD16+/CD56+ NK (p=0.007 and p<0.001, respectively), CD19+ (NS and p=0.001, respectively) and CD20+ cells (NS and p<0.001, respectively); there was no change in the percentage of HLA-DR+ T cells or effects of storage for 4 h. Effects of storage of blood in EDTA tubes at 4°C were limited to an increased percentage of CD8+ cells after 48 h (p<0.001) and a decreased percentage of CD4+ cells after 24 (p=0.009) and 48 h (p<0.001). When Streck Cell Preservative was added to the blood before storage for 24 or 48 h, changes in cell counts were limited to increases in the percentage of CD2+ cells (P=0.039 and P=0.005, respectively) and decreases in CD20+ cells (P=0.002 and P<0.0001, respectively). Interestingly, blood stored with Streck Cell Preservative had a higher percentage CD19+ (p<0.001) and lower percentage CD4+/CD3+ cells (P<0.001) after 4 h at room temperature, but these percentages were comparable to baseline when stored for 24 or 48 h.
The authors conclude that storage of either lithium heparin or EDTA blood for ≥24 h has a significant impact on lymphocyte subpopulations, but that while more subtypes are affected by storage of lithium heparin blood at 4°C than room temperature the lymphocyte population is more stable in EDTA blood at 4°C than room temperature. Importantly, the addition of Streck Cell Preservative to blood partially stabilized lymphocyte subpopulations.
Biospecimens
Preservative Types
- None (Fresh)
- Streck/BCT
Diagnoses:
- Normal
Platform:
Analyte Technology Platform Cell count/volume Flow cytometry Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Acquisition Type of collection container/solution EDTA tube
Lithium heparin tube
Biospecimen Acquisition Anticoagulant EDTA
Lithium heparin
Biospecimen Preservation Type of fixation/preservation Blood collection tube additive
EDTA
Refrigeration
None (fresh)
Storage Type of storage container EDTA tube
Lithium heparin tube
Tube with Streck Cell Preservative
Storage Storage duration 0 h
4 h
24 h
48 h
Storage Storage temperature 4°C
Room temperature