Pe mouse igg1

For cell surface markers analyses, cells were treated with 0.5% (w/v) bovine serum albumin and double-stained using monoclonal antibodies, including phycoerythrin (PE)-conjugated mouse anti-human CD34 (BD Biosciences, Le-Pont-de-Claix, France) with fluorescein isothiocyanate (FITC)-conjugated mouse anti-human CD90 (Beckman Coulter, Villepinte, France), PE-conjugated mouse anti-human CD166 (Beckman Coulter, Villepinte, France) with FITC-conjugated mouse anti-human CD45 (Dako, Glostrup, Denmark), PE-conjugated mouse anti-human CD105 (Beckman Coulter, Villepinte, France) with FITC-conjugated mouse anti-human CD44 (Beckman Coulter, Villepinte, France), and PE-conjugated mouse anti-human CD73 (BD Biosciences, Le-Pont-de-Claix, France) with FITC-conjugated mouse anti-human leukocyte antigen-antigen D related (HLA-DR) (Beckman Coulter, Villepinte, France). Antibodies were used at a dilution of 1:10 (PE CD34, FITC CD90, PE CD166, FITC CD44, PE CD105, FITC HLA-DR and PE CD73) or 1:20 (FITC CD45). Appropriate isotype-matched control antibodies named FITC or PE mouse IgG1 (Dako, Glostrup, Denmark) were used in each analysis. Cells were then examined by flow cytometry using a BD LSR II flow cytometer (Becton Dickinson Biosciences, San Jose, California). Fluorescence intensity and percentage of antigen positive cells were determined for each surface marker.

Cell surface marker expressions were assessed by
flow cytometry. The characterization panel consisted
of monoclonal antibodies for mesenchymal lineages
markers CD90-FITC (BD, Pharmingen^TM, USA), CD105PE
(Endoglin, BD Pharmingen^TM, USA), CD73-PE (BD
Pharmingen^TM, USA), CD44-FITC (BD Pharmingen^TM,
USA), CD45 FITC-CD34 PE (BD Pharmingen^TM, USA)
and CD11b (BD Pharmingen^TM, USA), along with the
following isotype controls, MultiMixTM FITC Mouse
IgG1, PE-Mouse IgG1 (Dako, Denmark), FITC-Mouse
IgG2b (Millipore, USA), and PE-conjugated Mouse
IgG1k (BD Pharmingen^TM, USA). Cells were fixed with
4% paraformaldehyde and immunophenotyping analysis
was performed by the BD FACS Calibur flow cytometry
system (BD Biosciences, USA).