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What Are Human Peripheral Blood Mononuclear Cells?

Human peripheral blood mononuclear cells (PBMCs) are a variety of immune cells derived from peripheral blood, and are defined as white blood cells with round nuclei. To achieve high yield and quality, PBMCs from �����ƽ�� are isolated from peripheral blood leukapheresis samples using density gradient separation and/ or red blood cell lysis (PBMCs can also be obtained from other, smaller sources such as whole blood, buffy coat, cord blood, and bone marrow). They are then cryopreserved in animal component-free CryoStor®CS10 medium (Catalog #07930). PBMCs are collected using ethically approved protocols, with Institutional Review Board (IRB) approval and in compliance with applicable legislation and guidance from the U.S. Food and Drug Administration (FDA) in the United States, or with Research Ethics Committee (REC) approval and in compliance with applicable legislation and guidance from the Human Tissue Authority (HTA) in the United Kingdom.

For a reference on the typical frequencies of each cell type in different cell sources, including PBMCs, leukopaks, and whole blood, get your free copy of the Frequencies of Human Cell Types in Blood-Related Sources wallchart.

PBMCs provide researchers with large cell volumes that can be used directly or processed further, offering flexibility for various therapeutic applications, including flow cytometry, cell isolation, cell culture, disease modeling, toxicology, and in vitro vaccine development. Collectively, these primary cells support cutting-edge advancements in cell and gene therapy research. Learn how PBMCs from �����ƽ�� can set your experiments up for success below.

Composition of Human Peripheral Blood Mononuclear Cells

PBMCs primarily include T lymphocytes (CD4⁺ and CD8⁺ subsets), B lymphocytes (CD19⁺), natural killer (NK) cells (CD16⁺/CD56⁺), and monocytes (CD14⁺) (Figure 1). These cells are typically isolated via Ficoll-Paque density gradient centrifugation or equivalent separation methods; please visit www.stemcell.com/pbmc for more information. The relative proportions of these subsets can vary based on donor characteristics such as age, sex, health status, and immune activation state. PBMCs are extensively used in immunological assays, including flow cytometry, ELISPOT, cytokine profiling, and functional cell-based assays, providing a critical ex vivo platform for modeling immune responses in preclinical and clinical research settings.

The largest population of cells in human PBMCs is made up of lymphocytes, of which T cells make up the majority, with B and NK cells comprising smaller proportions (Figure 2). These immune cells can be isolated and expanded, and are being used in current research for various applications.

Representative gating strategy of immune cell populations present in PBMCs. Flow cytometry was performed on the PBMCs post-thaw and can be provided for specific lots. The CD45 plot was gated on viable single cells while all other plots were gated on viable CD45+ single cells. In the above example, the cell frequencies are as follows: leukocytes (CD45+), 99.8%; B cells (CD19+), 10.5%; T Cells (CD3+), 57.2%; helper T cells (CD3+CD4+), 34.4%; cytotoxic T cells (CD3+CD8+), 19.6%; monocytes (CD14+), 17.9%; and NK cells (CD3-CD56+), 7.91%.

Applications of Human PBMCs in Preclinical and Clinical Research

• Immunotherapy
  PBMCs are commonly used during the development of immunotherapies, specifically for cytotoxicity and cytokine release assays, to demonstrate clinical safety, particularly regarding off-target effects and immunostimulatory potential. These PBMC-based assays provide crucial preclinical safety data that help mitigate risks before entering clinical trials. They are highly recommended to assess on-target/off-tumor effects, immunogenicity, and cytokine-mediated toxicities¹. In addition, using PBMCs from a diverse panel of donors (e.g. sex, different HLA types) helps predict variability in response, enhancing the translational relevance of results. PBMCs can also be profiled, before and after co-culture, by flow cytometry to observe activation markers on T cells, NK cells, or monocytes.
• Vaccine Development and Immunomonitoring
  PBMCs can be used in vitro to assess vaccine efficacy, safety, and to compare different vaccines. Specifically, PBMCs are used to evaluate T cell and B cell responses, and can bridge the gap between preclinical and clinical vaccine evaluation. Examples of vaccines where PBMCs have been used during development include some influenza² and Covid-19³ vaccines. PBMCs are also used to assess cell-mediated immunity, detect previous exposures to antigens, and monitor responses to immunotherapies.
• Immune Profiling in Disease Diagnosis and Monitoring
  Evaluating the proportions and activation status of PBMC types helps in understanding immune responses and monitoring various diseases, including autoimmune conditions, cancer, and infectious diseases. For example, in cancer, PBMCs can be used to assess the efficacy of immune checkpoint inhibitors and identify biomarkers related to cancer response and survival⁴.
• Development of Biologics and Small Molecules
  PBMCs provide a human-relevant in vitro system for screening biologics and small molecule drugs for immune-related effects. They are used to evaluate cytokine production, immune cell activation, potential immunogenicity, and cytotoxicity across a diverse donor pool. Key assays such as In Vitro Comparative Immunogenicity Assessment (IVCIA) help assess immunogenicity risk by comparing candidates to reference products. Multiplexed Analysis of Peptide-MHC Binding and T Cell Activation (MAPPs) enables detailed identification of potential T cell epitopes, improving immunogenicity prediction. These combined approaches help identify immune liabilities, support mechanism-of-action studies, and guide lead selection before entering the clinic.
• Assay Controls in Clinical Research
  PBMCs are designated for research use only, but are routinely applied in clinical research as reference samples in validated methods to monitor immune responses longitudinally, ensuring consistency and reliability across sites and timepoints. Healthy donor PBMCs act as baseline controls, with large donor numbers often required to represent diverse populations. These cells support potency, characterization, and release assays across biologics, cell and gene therapies, and vaccines. Recallable donors can be provided to support longitudinal studies and improve reproducibility, enabling standardized assay development and immune monitoring to support assay development, validation, and standardized immune monitoring throughout clinical development.

Other applications include: drug discovery screening and toxicity testing, transplantation and alloreactivity testing, infectious disease research, regenerative and anti-inflammatory therapies, and cell banking and biobanking.

Advantages of Frozen Human PBMCs

�����ƽ�� provides high-quality, highly characterized fresh (immediately after isolation) and frozen (cryopreserved for later use) PBMCs*. Frozen PBMCs offer flexibility and convenience because they can be stored long-term, and they also offer standardization when using the same cell populations across multiple experiments.

Have a question? See Frequently Asked Questions on Primary Cells for answers, or contact us directly.

Why Choose Human Peripheral Blood Mononuclear Cells from �����ƽ�� Technologies?

�����ƽ�� offers a comprehensive collection of ethically sourced, cryopreserved or fresh, diseased and normal PBMCs in different sizes to support your research needs (Table 2). In addition to the advantages listed in Table 1, �����ƽ�� provides further flexibility; mononuclear cells from peripheral blood are pre-aliquoted from a single donor, allowing researchers to purchase smaller vial quantities, pre-qualify donors or lots to ensure compatibility with their experiments, and place the remaining part of the lot on hold for future use.

�����ƽ�� has thousands of batches of HLA-typed (95% coverage) donors, with specific HLA types available on request, and hundreds available with flow cytometry data (standard panel; see Figure 2), so you can rely on proven quality and consistency. Further features of PBMCs available as custom requests include: specific donor demographics, additional viral testing, additional genotyping, additional donor information or medical history (medication, etc). �����ƽ�� also has recallable donors and different sizes available to suit your research needs. Find out more about custom solutions here.