Anti cd90 pe cy7 clone 5e10

Multiparameter analysis of purified cells for surface antigen expression was performed by incubating thawed or cultured cells at room temperature for 15 minutes in PBS/1% bovine serum albumin and washed twice before flow cytometric analysis. The monoclonal antibodies were anti-CD34 APC-Cy7 (clone 581; Biolegend, USA), anti-CD38 V450 (clone HIT2; Exbio, Czech Republic), anti-CD90 PE-Cy7 (clone 5E10; Biolegend), anti-CD47 PerCP Cy5.5 (clone CC2C6; Biolegend), PerCP Cy5.5 isotype (clone MOPC-21, Biolegend), and lineage cocktail FITC against CD3, CD14, CD16, CD19, CD20, CD56 (Cat.No.6K01-T050; Exbio). Flow cytometric analysis was performed on a Beckton-Dickson FACS Canto II. A minimum of 2400 events (median 16,500) for CD34 positive cells were obtained in each sample.
For CD47 expression analysis of non-cultured cells, we defined different cell populations as follows: HSC (CD34+CD38- CD90+Lin-), pluripotential progenitor cells (PPC) (CD34+CD38-CD90-Lin-), lineage-committed progenitors (LCP) (CD34+CD38+Lin-), and differentiated cells (DC) (CD34-Lin+) of patients and controls. Figure 1 shows the gating strategy. For every population, CD47 expression intensity was calculated by subtracting its mean fluorescence intensity (MFI) from the isotype MFI.