A total of 100 μL of blood, or 1–2 × 105 PBMCs or 0.5–1 × 105 culture cells, removed from the plates and washed with PBS were taken for staining. Сombined staining was performed with conjugated monoclonal antibodies against human antigens produced by Biolegend (San Diego, CA, USA) (unless otherwise specified): fluorescein isothiocyanate (FITC)—(CD3, CD34, vWF (Abcam, Cambridge, UK), phycoerythrin (PE)—(KDR (BD), CD14), allocicocyanin (APC)—(CD133, CD31), phycoerythrin-cyanine 7 (PC7) CD146, PacificBlue 450 (PB 450) HLA DR, Krome Orange (KrOr) CD45 (BC, Beckman Coulter, Brea, CA).
Whole PB, PBMCs, and cell cultures were allocated according to the manufacturer’s protocols in two panels:
Blood sample preparation additionally included the lysing of red blood cells with VersaLyse (BC, Beckman Coulter, Brea, CA, USA) and washing with PBS. Fixation and permeabilization of the cells were performed using the IntraPrep (BC) kit when staining intracellular proteins with vWF. The stained samples were resuspended in PBS and analyzed on a CytoFlex flow cytometer (USA) with CytExpert software (version 2.1, Beckman Coulter, CA, USA).
Whole PB, PBMCs, and cell cultures were allocated according to the manufacturer’s protocols in two panels:
CD3, CD14, HLADR, CD45
CD34, KDR, CD146, CD133, CD31, CD45
Blood sample preparation additionally included the lysing of red blood cells with VersaLyse (BC, Beckman Coulter, Brea, CA, USA) and washing with PBS. Fixation and permeabilization of the cells were performed using the IntraPrep (BC) kit when staining intracellular proteins with vWF. The stained samples were resuspended in PBS and analyzed on a CytoFlex flow cytometer (USA) with CytExpert software (version 2.1, Beckman Coulter, CA, USA).