*2.12. Flow Cytometry*

For immunophenotyping of MSCs, adhesive cells of the 2nd-3rd passages were collected using Accutase (CLS, Eppelheim, Germany), washed twice in ice-cold PBS, and separately stained (106 cells/sample) with primary phycoerythrin (PE)-labeled antibodies against CD73, CD90.1, and CD105 (BD Biosciences, USA) for 45 min, or unlabeled rat antibodies against CD45 and CD34 for 45 min and secondary Rabbit Anti-Rat-FITC conjugate (Abcam, Cambridge, UK) for 30 min at room temperature.

Analysis of dendritic (DC) and myeloid derived suppressor cells (MDSCs) was performed by multicolor flow cytometry. The suspensions of splenocytes from 10 immunized mice of each group

(106 cells/sample) were incubated with PE-labeled antibodies against CD11c, FITC-labeled antibodies against Gr-1 (Ly-6G and Ly-6C), and allophycocyanin (APC)-labeled antibodies against CD11b (BD Biosciences, USA). Antibodies to the corresponding isotype controls were included in all experiments. The absolute and relative number of cells carrying the markers was assessed by FACS on a flow cytometer BD FACSCanto II (Beckton Dickinson, Franklin Lakes, NJ, USA) using free software BD FACSDiva, v.6.1.3 (BD Biosciences, San Jose, CA, USA).
