Separation and purification of MDSC in vitro
BALB/c mice were sacrificed, then bone marrow cells were extracted and made into single cell suspension. After red blood cells having been cracked, cell suspension was administrated by anti-Gr-1 monoclonal Ab (mAb), anti-CD11b mAb, anti-CD80 mAb, anti-F4/80 mAb, anti-CD11c mAb and anti-MHCII mAb. Differentiation of MDSC and proportion of Gr-1+CD11b+MDSCs were detected by flow cytometry.
Bone marrow cells were suspended by PBS and MDSC were separated by Percoll Density Gradient Centrifugation. Then, cell suspension was collected and separated by CD11b magnetic bead and purity of Gr-1+CD11b+MDSC was detected by flow cytometry.