Biospecimen Science of Blood for Peripheral Blood Mononuclear Cell (PBMC) Functional Applications

Abstract

Purpose of Review Peripheral blood mononuclear cells (PBMCs) are used in a wide variety of preclinical assays. Preanalytical variations can have a devastating impact on the results. In this review, we list critical preanalytical factors for PBMC-based assays to develop awareness and orientation to the types of sample preparation and storage that one may consider employing. Recent Findings Critical factors during blood collection are the blood collection tube and anticoagulant, possible stabilizer used, and the pre-isolation blood storage temperature and time. During PBMC isolation, critical factors are the isolation method, density gradient or magnetic sorting, use of barrier, possible RBC lysis, and centrifuge type. During cryopreservation, attention is needed for the cryomedium type and temperature, freezing device and program, cell concentration, and the long-term storage temperature. During the thawing process, the thawing procedure/device used and wash medium temperature are critical. Summary To avoid biased results in PBMC assays, these critical preanalytical factors must be standardized and/or documented. Additionally, participation in external quality assurance programs is strongly recommended.

The Cytokinesis-Block Micronucleus Assay on Human Isolated Fresh and Cryopreserved Peripheral Blood Mononuclear Cells

“…While WBC are commonly used for cytogenetic assays, isolated PBMCS are also finding their way in chromosomal damage research [21][22][23]. The potential for cryopreservation of PBMCS makes this an exceptionally suitable cell type for cytogenetic assays, as it has several major advantages: (1) the ability to obtain a bulk of cells; (2) no need for further blood sampling; (3) the potential for batching and (4) the ability to perform the analysis within a chosen time frame.…”

“…Furthermore, as stated by various research groups, care should be taken regarding differences in isolation protocols and storage conditions for all PBMC samples in one study. In addition, it is important to provide a precise description of preanalytical processing and variations [23,32].…”

“…All pre-analytical factors used for PBMC-based assays are documented according to the recent standards determined by Betsou et al [23].…”

Streptococcus thermophilus alters the expression of genes associated with innate and adaptive immunity in human peripheral blood mononuclear cells

“…Calcium and magnesium free PBS, pH7.2, (Invitrogen, Pty Ltd. Australia) was used after adding 2 mM EDTA and 2% heat-inactivated fetal bovine serum (FBS) (Invitrogen, Pty Ltd. Australia); PBS buffer. SEPMATE tubes (50 ml) with inner inserts (STEMCELL technology, Canada) were used to isolate PBMC following Ficoll-Paque density gradient protocol [29,30]. PBMCs were washed, counted and the required number of PBMC were co-cultured with S. thermophilus 285 and the remaining PBMC were stored in freeze mix and transferred into liquid nitrogen for future use.…”

Role of the CXCR4-SDF1-HMGB1 pathway in the directional migration of cells and regeneration of affected organs

“…PBMC are widely used in preclinical research and applications in vaccine trials, source of biomarkers in various infectious and chronic diseases, and are a useful tool in studying various aspects of pathology and biology in vitro [ 15 ]. In addition, PBMC are an easily accessible source of different types of adult stem and progenitor cells, such as HSCs, MSCs, osteoclast precursor cells, and EPCs[ 16 ].…”