Heparinized fresh whole blood was obtained from healthy donors of the NIH Clinical Center Department of Transfusion Medicine (n= 32) after informed consent was obtained in accordance with the Declaration of Helsinki [32 (link)] (NIH, Bethesda, MD, USA). Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll-Paque gradient centrifugation (GE Healthcare, Chicago, IL) with Leucosep centrifuge tubes (VWR, Radnor, PA), according to manufacturer’s instructions. Cells were frozen in medium containing 90% FCS and 10% dymethyl-sulfoxide (DMSO, Sigma-Aldrich, St. Louis, MO, USA) and stored in liquid nitrogen until use.
Leucosep centrifuge tubes
Manufactured by Avantor
Leucosep centrifuge tubes are a type of laboratory equipment used for the separation and isolation of cells, particularly white blood cells (leukocytes), during centrifugation. These tubes feature a porous barrier that allows for the efficient separation of the desired cell population from the rest of the sample.
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Lab products found in correlation
2 protocols using leucosep centrifuge tubes
1
PBMC Isolation and Cryopreservation
2
PBMC Isolation from Whole Blood
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Blood from healthy donors was collected in EDTA tubes and PBMCs were isolated using Leucosep centrifuge tubes (VWR) following the manufacturer's instructions. Briefly, the bottom chamber was filled with Lymphoprep and top chamber overlaid with fresh patient whole blood. Tubes were spun at 1000g for 10 min, and the buffy coat was collected. Cells were washed with RPMI 1640 media containing 10% FBS, 100 U/mL penicillin, 100 μg/mL streptomycin, and 1% Glutamax. For later use, cells were stored in vapor phase liquid nitrogen in freezing media containing 10% DMSO.
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