ArticleBMC immunology2025
Effect of delayed isolation of peripheral blood mononuclear cells on cell viability and functionality.
Article in BMC immunology, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
5 citing papers in PubMed.
- Review
- Integrated clinical and PBMC transcriptomic profiling identifies lipid metabolism-related candidate signatures associated with chronic brucellosis.Frontiers in cellular and infection microbiology · 2026Article
- Refrigerated storage of L-PRF membranes: anti-bacterial activity, cytokine release, andFrontiers in oral health · 2026Article
- Comprehensive real-time metabolic profiling of peripheral blood mononuclear cells reveals important methodological considerations for immunometabolism research.Frontiers in immunology · 2025Article
- My cells, my model: immune-competent autologous organ-on-chip systems as a new paradigm in precision medicine.Frontiers in immunology · 2025Review
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Authors and funding
12 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundPeripheral blood mononuclear cells (PBMCs) are valuable biomarkers, providing crucial insights into the patients' immune system. Reliable biobanking of PBMCs is essential to minimize heterogeneity. In multicenter trials, blood sample transportation to central laboratories can increase the time between blood collection and PBMC isolation. This study evaluated the effect of prolonged blood hold time on PBMC viability and cytotoxicity.
methodsFrom July 2021 to May 2023, 104 patients with early HER2-positive breast cancer were enrolled in the NeoOn trial, of whom 49 patients were included in this subproject. PBMCs were isolated ≤ 6 hours (h) or ≥ 20 h after blood collection. PBMC yield and viability were determined using the LUNA-II Automated Cell Counter. Flow cytometry was used to quantify in vitro cytotoxicity, the percentage of natural killer (NK) and T cells, as well as apoptotic and necrotic cells.
resultsIsolating PBMCs ≥ 20 h resulted in a higher cell yield, but lower NK cell viability compared to PBMCs ≤ 6 h. PBMCs ≥ 20 h were less robust to thawing and showed higher loss during recovery. Compared to PMBCS ≤ 6 h, PBMCs ≥ 20 h exhibited lower antibody-mediated cytotoxicity (p ≤ 0.0001) and antibody-dependent phagocytosis (p < 0.0051). While the percentage of T and NK cells and the T cell viability remained unaffected by hold time, the percentage of apoptotic NK cells was higher for PBMCs ≥ 20 h (41.0 ± 12.9% vs. 23.8 ± 13.4%; p = 0.0364).
conclusionsExtended blood storage time caused increased apoptosis and necrosis of NK cells, adversely affecting PBMC quality and reducing NK cell related functionality. Hence, blood hold time should be minimized to maintain PBMC integrity and NK cell functionality for in vitro biomarker assays.
trial registrationTrial registration number: EudraCT 2020-001943-21. Date of registration: December 29th 2020.
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