Isolation and freezing of human peripheral blood mononuclear cells from pregnant patients

• Published in: STAR protocols Vol. 3; no. 1; p. 101204
• Main Authors: Efthymiou, Athina, Mureanu, Nicoleta, Pemberton, Rebecca, Tai-MacArthur, Sarah, Mastronicola, Daniela, Scottà, Cristiano, Lombardi, Giovanna, Nicolaides, Kypros H., Shangaris, Panicos
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 18.03.2022; Elsevier
• Subjects: CD8-Positive T-Lymphocytes; Cell Biology; Cell isolation; Clinical Protocol; Female; Freezing; Health Sciences; Humans; Immunology; Leukocytes; Leukocytes, Mononuclear; Pregnancy; Protocol; Stem Cells; T-Lymphocytes, Regulatory; Stem Cells; Immunology; Clinical Protocol; Cell isolation; Cell Biology; Health Sciences
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2022.101204

To analyze immune cell populations accurately, a large number of Peripheral Blood Mononuclear Cells (PBMCs) must be obtained from blood samples. Traditional manual isolation and SepMate™ isolation of PBMCs consistently yield blood-stained plasma layers and overall low numbers of CD4+ and CD8+ cells. Here, we describe an optimized protocol, using PBS with EDTA to increase PBMC yield from pregnant patients. This protocol enables analysis of CD4+, CD8+, and Regulatory T Cells and is potentially applicable to any immune cell population. For complete details on the use and execution of this protocol, please refer to the SepMateTM website https://www.stemcell.com/products/brands/SepMateTM-pbmc-isolation.html. [Display omitted] •SepMate™ Tubes offer fast and efficient PBMC isolation•Using PBS with 1 mM EDTA increases the yield from pregnant patients’ blood samples•Careful freezing ensures increased viability of cells•Achieves increased viability by following a step-by-step thawing process To analyze immune cell populations accurately, a large number of Peripheral Blood Mononuclear Cells (PBMCs) must be obtained from blood samples. Traditional manual isolation and SepMate™ isolation of PBMCs consistently yield blood-stained plasma layers and overall low numbers of CD4+ and CD8+ cells. Here, we describe an optimized protocol, using PBS with EDTA to increase PBMC yield from pregnant patients. This protocol enables analysis of CD4+, CD8+, and Regulatory T Cells and is potentially applicable to any immune cell population.