Ficoll paque plus kit

Samples of peripheral blood from 10 healthy people who consciously and voluntarily submitted them for testing and signed individual statements were collected after obtaining the consent of the Bioethics Committee at the Medical University of Wroclaw, No. 71/2017. First, whole blood was collected into test tubes with EDTA anticoagulant by specialized personnel of the Diagnostics laboratory at the Institute of Immunology and Experimental Therapy. According to the manufacturer’s instructions, blood mononuclear cells were then isolated using the Ficoll-Paque PLUS kit (GE Healthcare, Amersham, UK).

Peripheral blood mononuclear cells (PBMCs) were isolated by gradient centrifugation using a Ficoll-Paque PLUS kit (GE Healthcare Life Sciences, Pittsburgh, PA, United States). Briefly, the blood was diluted with the same volume of phosphate buffered saline (PBS) (Solarbio, Beijing, China). The diluted samples were carefully layered onto the Ficoll-Paque medium solution, and then centrifuged at 400 × g for 35 min at 20°C. Afterward, the mononuclear cell layers were transferred into new tubes and washed with PBS twice. The brake of the centrifuge was turned off to minimize disturb of the mononuclear layer.

Peripheral blood samples (10 mL each) from 10 healthy people who consciously and voluntarily submitted them for testing and signed individual statements were collected after obtaining the consent of the Bioethics Committee at the Medical University of Wroclaw, No. 71/2017. Whole blood was collected in test tubes with an EDTA anticoagulant by specialized personnel of the diagnostics laboratory at the Institute of Immunology and Experimental Therapy. Blood mononuclear cells were then isolated using the Ficoll-Paque PLUS kit (GE Healthcare, Amersham, UK) according to the manufacturer’s instructions.