Phycoerythrin conjugated antibodies against human cd73 and cd166

For the cytometric analysis of cultured cell phenotypes, the cells were stained for 15 min at room temperature with fluorescein isothiocyanate-conjugated antibodies against human CD14, CD45, and HLA-DR (BD Biosciences); phycoerythrin-conjugated antibodies against human CD73 and CD166 (BD Biosciences); and CD105 (Serotec, Kidlington, UK). Corresponding isotype-matched mouse antibodies were used as controls. The cells were washed with PBS and fixed with 1% (v/v) paraformaldehyde (Sigma-Aldrich). The immunotype of the MSCs was determined by flow cytometry on a FACSCalibur instrument, and then the percentage of expressed cell surface antigens was calculated for 10,000 gated cell events.

For cytometric analysis of the cultured cell phenotypes, the cells were stained for 15 min at room temperature with fluorescein isothiocyanate-conjugated antibodies against human CD14 and CD45 and human leukocyte antigen-DR isotype (BD Biosciences, Franklin Lakes, NJ, USA); phycoerythrin-conjugated antibodies against human CD73 and CD166 (BD Biosciences) and CD90 and CD105 (Invitrogen). Corresponding isotype-matched mouse antibodies were used as controls. The cells were washed with phosphate-buffered saline (PBS, Corning, Manassas, VA, USA) and fixed with 1% (v/v) paraformaldehyde (Sigma-Aldrich). The immunotype of the MSCs was determined by flow cytometry on a FACSCalibur instrument (BD Biosciences), and the percentage of expressed cell surface antigens was calculated for 10,000 gated-cell events [26 (link)].