ArticlePloS one2017
Optimizing recovery of frozen human peripheral blood mononuclear cells for flow cytometry.
Article in PloS one, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed.
- Effect of delayed cell processing and cryopreservation on immunophenotyping in multicenter population studies.Journal of immunological methods · 2018Trial
- Mitochondrial dysfunction and autophagy activation underlie NK cell impairment induced by Cannabis.PloS one · 2026Article
- Porcine Peripheral Blood Mononuclear Cells (PBMCs): Methods of Isolation, Cryopreservation, and Translational Applications in Human Studies.Journal of clinical medicine · 2025Review
- Bioenergetic and immunological characterization of cryopreserved peripheral blood mononuclear cells (PBMCs) isolated from blood and buffy coat.Frontiers in molecular biosciences · 2025Article
- Efficacy of Quality and Quantity media-cultured mononuclear cells for promoting peripheral nerve regeneration in mouse model.PloS one · 2025Article
- Improving Reliability of Immunological Assays by Defining Minimal Criteria for Cell Fitness.ImmunoHorizons · 2024Article
- Humoral and Innate Immunological Profile of Paediatric Recipients of Pfizer-BioNTech BNT162b2 mRNA Vaccine.Microorganisms · 2024Article
- Preservation of functionality, immunophenotype, and recovery of HIV RNA from PBMCs cryopreserved for more than 20 years.Frontiers in immunology · 2024Article
- Research progress on optimization ofFrontiers in bioengineering and biotechnology · 2024Review
- Glycolysis Changes in Alloreactive Memory B Cells in Highly Sensitized Kidney Transplant Recipients Undergoing Desensitization Therapy.Transplant international : official journal of the European Society for Organ Transplantation · 2024Article
- scRNA-seq reveals elevated interferon responses and TNF-α signaling via NFkB in monocytes in children with uncomplicated malaria.Experimental biology and medicine (Maywood, N.J.) · 2024Article
- Technical pitfalls when collecting, cryopreserving, thawing, and stimulating human T-cells.Frontiers in immunology · 2024Review
- Standardized assays to monitor drug sensitivity in hematologic cancers.Cell death discovery · 2023Article
- Anti-Inflammatory Effects of Nutritionally Relevant Concentrations of Oleuropein and Hydroxytyrosol on Peripheral Blood Mononuclear Cells: An Age-Related Analysis.International journal of molecular sciences · 2023Article
- High-throughput measurement of double strand break global repair phenotype in peripheral blood mononuclear cells after long-term cryopreservation.Cytometry. Part A : the journal of the International Society for Analytical Cytology · 2023Article
- Qualification of Hemophilia Treatment Centers to Enable Multi-Center Studies of Gene Expression Signatures in Blood Cells from Pediatric Patients.Journal of clinical medicine · 2023Article
- The quality and quantity media-cultured mononuclear cell transplantation is safe and effective in ischemic hindlimb mouse model.JVS-vascular science · 2023Article
- Quantifying the Detrimental Effects of Multiple Freeze/Thaw Cycles on Primary Human Lymphocyte Survival and Function.International journal of molecular sciences · 2022Article
- Immune Subsets From Ficoll Density Gradient Separation in Kidney Transplant Recipients.Transplantation direct · 2022Article
- Novel culture media enhances mononuclear cells from patients with chronic limb-threatening ischemia to increase vasculogenesis and anti-inflammatory effect.Stem cell research & therapy · 2021Article
Corrections and comments
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
introductionLive peripheral blood mononuclear cells (PBMCs) can be frozen and thawed for later analyses by adding and removing a cryoprotectant, such as dimethyl sulfoxide (DMSO). Laboratories across the world use various procedures, but published evidence of optimal thawing procedures is scarce. MATERIALS AND
methodsPBMCs were separated from blood collected from healthy Danish blood donors, and stored at -80°C after adding of DMSO. The essential steps in the thawing procedure were modified and performance was evaluated by flow cytometry with respect to the percentage and total yield of viable PMBCs.
resultsThe best-performing washing medium was Roswell Park Memorial Institute (RPMI) 1640 at 37°C with 20% fetal bovine serum. When using 10 mL washing medium in a 15-mL Falcon tube, samples should be centrifuged for at least 10 minutes at 500 g. We failed to detect any differences between the tested methods of mixing PBMCs with washing medium. Likewise, neither the thawing duration nor centrifugation temperature (20°C and 37°C) had any effect. PBMCs could be incubated (rested) for up to eight hours in a 37°C 5% CO2 incubator without affecting cell counts, but incubating PBMCs for 16 hours significantly decreased viability and recovery. In general, high viability was not necessarily associated with high recovery.
conclusionChanging the thawing procedure significantly impacted PBMC viability and live cell recovery. Evaluating both viability and live PBMC recovery are necessary to evaluate method performance. Investigation of differential loss of PBMC subtypes and phenotypic changes during thawing and incubation requires further evaluation.
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