Peripheral Blood Mononuclear Cell Isolation

A total of 15 mL of peripheral blood was collected in EDTA-coated tubes (Sarstedt, Nümbrecht, Germany), and PBMCs and plasma were isolated by density gradient centrifugation (using Ficoll-Paque™; Milteny Biotec, Bergisch-Gladbach, Germany) at 400 g for 30 min. Plasma was stored at −80 °C until use. Only PBMCs with a viability of ≥95% (determined using 0.4% trypan blue solution; Sigma Aldrich, Hamburg, Germany) were used in subsequent experiments [25 (link),30 (link)].

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Grywalska E., Smarz-Widelska I., Korona-Głowniak I., Mertowski S., Gosik K., Hymos A., Ludian J., Niedźwiedzka-Rystwej P., Roliński J, & Załuska W. (2020). PD-1 and PD-L1 Expression on Circulating Lymphocytes as a Marker of Epstein-Barr Virus Reactivation-Associated Proliferative Glomerulonephritis. International Journal of Molecular Sciences, 21(21), 8001.

Publication 2020

EDTA-coated tubes Ficoll-Paque trypan blue solution

Blood Density gradient centrifugation Edta Ficoll Plasma Trypan blue

Corresponding Organization : Medical University of Lublin

Other organizations : University of Szczecin

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Variable analysis

independent variables

Density gradient centrifugation (using Ficoll-Paque™; Milteny Biotec, Bergisch-Gladbach, Germany) at 400 g for 30 min

dependent variables

PBMC viability (determined using 0.4% trypan blue solution; Sigma Aldrich, Hamburg, Germany)

control variables

Total volume of peripheral blood collected (15 mL)
EDTA-coated tubes (Sarstedt, Nümbrecht, Germany) used for blood collection
Plasma stored at -80 °C until use

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