Porcine Peripheral Blood Mononuclear Cells (PBMCs): Methods of Isolation, Cryopreservation, and Translational Applications in Human Studies

Abstract

Porcine peripheral blood mononuclear cells (pPBMCs) are increasingly recognized as a valuable model in biomedical and translational research, particularly in contexts directly related to human health and disease. Their immunological features, such as the presence of CD4⁺CD8⁺ double-positive T cells and cytokine expression patterns, exhibit a notable degree of similarity to human immune cells, making them an attractive tool for studying human-relevant immune responses. This review outlines current methodologies for isolating and cryopreserving pPBMCs, with a focus on maintaining high cell viability and functionality. Key technical considerations, including the optimal use of gradient media, appropriate anticoagulants, and standardized freezing/thawing protocols, are discussed in detail. Furthermore, the article highlights the applications of pPBMCs in various research contexts, including vaccine development, inflammation studies, infection models, and xenotransplantation. A comparative perspective is provided to identify similarities and differences between porcine and human PBMCs, supporting the validity of swine as a translational model. Evidence from pPBMC-based studies has shown predictive value for human outcomes, reinforcing their role as a surrogate system for preclinical investigations. Given their anatomical, physiological, and immunogenetic similarities to humans, porcine PBMCs represent a valuable bridge between basic science and clinical application, playing an increasingly important role in translational medicine.

Keywords: PBMC; cryopreservation; isolation; peripheral blood mononuclear cells; pig; porcine; swine model.

Figure 3

Comparison of the most characteristic markers for porcine and human PBMCs. The bracket indicates that the identification of these cells requires the use of multiple markers, which allows for their accurate differentiation. Abbreviations: B = B lymphocyte; CD = cluster of differentiation; FoxP3 = Forkhead box protein P3; γδ T = gamma–delta T lymphocyte; mDC = myeloid dendritic cell; NK = natural killer cell; pDC = plasmacytoid dendritic cell; PBMC = peripheral blood mononuclear cells; T = T lymphocyte; Tc = cytotoxic T lymphocyte; TCR = T cell receptor; Th = T helper lymphocyte; Treg = regulatory T lymphocyte [39,49,50,51,52,53,54,55,56,57,58].

Figure 1

Protocol of PBMC isolation from peripheral blood of swine. Abbreviations: PBMC = peripheral blood mononuclear cells; PBS = phosphate-buffered saline; RBC = red blood cells; RPMI 1640 = Roswell Park Memorial Institute 1640 medium.

Figure 2

Comparison of blood sample separation using 50 mL tube and SepMate™-50 tube. Abbreviations: B = B lymphocyte; Bas = basophils; DC = dendritic cells; Eos = eosinophils; Mo = monocytes; NK = natural killer cells; Neu = neutrophils; PBMC = peripheral blood mononuclear cells; RBC = red blood cells; T = T lymphocyte.