The work with peripheral blood mononuclear cells (PBMCs), which comprise lymphocytes and monocytes, is indispensable in immunological diagnostics and research. The isolation of PBMCs takes advantage of differences in cell density of the different blood components. Density gradient centrifugation of diluted whole blood layered over a density gradient medium yields PBMCs; two subsequent washing steps remove remaining platelets. To store the cells for future assays, they can be frozen and thawed when required. Dimethyl sulfoxide (DMSO) serves as a cryoprotectant for freezing PBMCs, but must be removed by washing after thawing, as it can become toxic to the cells on longer exposure.
PBMC Density gradient Freezing Thawing Cryopreservation
This is a preview of subscription content, log in to check access.
Springer Nature is developing a new tool to find and evaluate Protocols. Learn more
Wang SY, Hsu ML, Tzeng CH et al (1998) The influence of cryopreservation on cytokine production by human T lymphocytes. Cryobiology 37:22–29PubMedCrossRefGoogle Scholar
Kreher CR, Dittrich MT, Guerkov R et al (2003) CD4+ and CD8+ cells in cryopreserved human PBMC maintain full functionality in cytokine ELISPOT assays. J Immunol Methods 278:79–93PubMedCrossRefGoogle Scholar
Weinberg A, Song L, Wilkening C et al (2009) Optimization and Limitations of Use of Cryopreserved Peripheral Blood Mononuclear Cells for Functional and Phenotypic T-Cell Characterization. Clin Vaccine Immunol 16:1176–1186PubMedCentralPubMedCrossRefGoogle Scholar
Costantini A, Mancini S, Giuliodoro S et al (2003) Effects of cryopreservation on lymphocyte immunophenotype and function. J Immunol Methods 278:145–155PubMedCrossRefGoogle Scholar
Bull M, Lee D, Stucky J et al (2007) Defining blood processing parameters for optimal detection of cryopreserved antigen-specific responses for HIV vaccine trials. J Immunol Methods 322:57–69PubMedCentralPubMedCrossRefGoogle Scholar
Ramachandran H, Laux J, Moldovan I et al (2012) Optimal Thawing of Cryopreserved Peripheral Blood Mononuclear Cells for Use in High-Throughput Human Immune Monitoring Studies. Cells 1:313–324PubMedCentralPubMedCrossRefGoogle Scholar
Disis ML, dela Rosa C, Goodell V et al (2006) Maximizing the retention of antigen specific lymphocyte function after cryopreservation. J Immunol Methods 308:13–18PubMedCrossRefGoogle Scholar
Kloverpris H, Fomsgaard A, Handley A et al (2010) Dimethyl sulfoxide (DMSO) exposure to human peripheral blood mononuclear cells (PBMCs) abolish T cell responses only in high concentrations and following coincubation for more than two hours. J Immunol Methods 356:70–78PubMedCrossRefGoogle Scholar