Apc conjugated cd105

iMSCs (iPSCs-derived MSCs) were suspended at a concentration of 1 × 10⁵ cells/mL in 1 × DPBS. A total of 5 × 10⁴ cells were incubated with BB515-conjugated CD44, Precp-Cy5.5-conjugated CD73, PE-Cy7-conjugated anti-human CD90, APC-conjugated CD105, BV421-conjugated anti-human CD34, CD45, and HLA-DR (BD Biosciences) at room temperature for 30 min in the dark. Then, the stained cells were washed twice in DPBS. Flow cytometry analysis was performed by a flow cytometer (BD Biosciences) to detect the expression of the cell surface molecules of differentiated iMSCs.

The iMSCs were dissociated into single cells with TrypLE™ Express and counted, and 5 × 10⁴ cells were suspended in 100 μL DPBS with 5% BSA. Then cells were incubated with BB515-conjugated CD44, Precp-Cy5.5-conjugated CD73, PE-Cy7-conjugated anti-human CD90, APC-conjugated CD105, BV421-conjugated anti-human CD34, CD45, and HLA-DR (BD Biosciences, Franklin Lakes, NJ, USA) at 4 °C for 30 min in the dark. The stained cells were washed twice in DPBS and characterized by flow cytometry (BD Biosciences, Franklin Lakes, NJ, USA).
The differentiation potential of iMSCs was identified via osteogenesis (Gibco, New York, NY, USA), adipogenesis (STEMCELL Technologies, Vancouver, BC, Canada), and chondrogenesis (STEMCELL Technologies, Vancouver, BC, Canada) differentiation kits, according to the manufacturer’s instructions. After being cultured with differentiation medium for 2 to 3 weeks, cells were stained with alizarin red, oil red O, or alcian blue dye (Cyagen Biosciences Inc., Guangzhou, China)) for 30 min separately, then the stained cells were analyzed using a light microscope.

The cell suspension was prepared at a concentration of 1 × 10⁵/mL in 1 × DPBS. 5 × 10⁴ cells were incubated with BV421-conjugated anti-human CD34, CD45 and HLA-DR, BB515-conjugated CD44,Precp-Cy5.5-conjugated CD73, APC-conjugated CD105 and PE-Cy7-conjugated anti-human CD90 (BD Biosciences, USA) at room temperature for 30 min. Stained cells were then washed twice in PBS. Flow cytometric analysis was performed by flow cytometer (BD Biosciences, USA) to detect the expression of cell surface markers of iMSCs and IL-24-iMSCs.