Cd90 r pe

The osteogenicity of MSCs from the orbicularis oris muscle has been studied by our laboratory and has been previously shown to induce in vitro and in vivo bone formation in critical-sized calvarial osseous defects in rats.^{20 (link)}Flow cytometry analysis was performed using a Guava Easy Cyte microcapillary flow cytometer (Guava Technologies, Hayward, CA, USA) utilizing laser excitation and emission wavelengths of 488 and 532 nm. Cells were pelleted and resuspended in phosphate-buffered saline (PBS; Gibco-Invitrogen^®, Carlsbad, CA, USA) at a concentration of 1.0 × 10⁶ cells/mL and stained with saturating concentrations of antibodies. After a 45-min incubation in the dark at room temperature, cells were washed three times with PBS and resuspended in 0.25 mL of cold PBS. In order to analyze cell surface expression of typical protein markers, adherent cells were treated with the following primary anti-human antibodies: CD29-PE-Cy5, CD31-PE, CD45-FITC, CD73-PE, CD90-R-PE, and CD105-PE (Becton Dickinson, Franklin Lakes, NJ, USA). Unstained cells were gated on forward scatter to eliminate particulate debris and clumped cells. A minimum of 5000 events were counted for each sample.
These cells presented high expression of MSC markers (CD73-PE, CD90-R-PE, CD105-PE) and lack of expression of hematopoietic and endothelial markers (CD29-PE-Cy5, CD31-PE, CD45-FITC).