Fitc conjugated nonspecific immunoglobulin g igg

At passage 0, the characteristics of younger and older MSCs from clotted BMAs were investigated by characterizing cell-surface markers of isolated cells using flow cytometry analysis. To test surface antigen expression, 0.5–1 × 10⁵ MSCs for each antigen was washed with PBS, centrifuged at 260 g for 5 min, and incubated at 4 C for 30 min in flow cytometry buffer (FCB, 2% FBS in PBS) adding 0.5 μg/ml of fluorescein isothiocyanate (FITC)-conjugated antibody against CD31, CD45, CD34, CD44, CD73, CD90, and CD105. FITC-conjugated nonspecific immunoglobulin G (IgG) was used as isotype control (BioLegend, San Diego, CA, United States). Cell fluorescence was evaluated with FACSCanto II instrument (Becton Dickinson, Franklin Lakes, NJ, United States) and analyzed by FACSDiva software (Becton Dickinson).

Antigen expression was performed as previously described [14 (link)] and assessed with FACSCanto II instrument (Becton Dickinson, Franklin Lakes, NJ, USA) and by FACSDiva software 6.0 (Becton Dickinson). Briefly, at passage 1, 0.5–1 × 10⁵ of MSCs for each antigen were washed with PBS, centrifuged, and incubated in flow cytometry buffer, adding fluorescein isothiocyanate (FITC)-conjugated antibody against CD-31, 45, 34, 44, 73, 90, and 105. FITC-conjugated nonspecific immunoglobulin G (IgG) was used as control (Bio-Legend, San Diego, CA, USA).