Cd274 pe

hWJ-MSCs were immunophenotypically characterized by labeling the cells with anti-CD90-APC, -CD73-PE/Cy7, -CD105-PE, -CD274-PE, -CD45-APC/Cy7, -CD34-PerCP- Cy5.5, and -CD31-PE antibodies (Biolegend, San Diego, USA). T-cell depletion lymphocytes (TCD3⁺) inhibition was evaluated and cell suspensions were analyzed for sterility, endotoxins, and mycoplasma. A FACSCanto II cytometer (BD, Franklin Lakes, NJ, USA) and FlowJo vX.7.0 software (TreeStar, USA) was used for flow cytometry and data analysis, respectively.

Expanded WJ-MSC (n = 3) supplemented with 10% hPL (n = 3, batch 50, 53, 54), hS (n = 3, batch 21, 54, 57) and FBS (n = 2, batch 1, 2) (Gibco, Life Technologies, Carlsbad, CA, USA) were characterized immunophenotypically using antibodies against the following human antigens—CD90-APC, CD73-PE/Cy7, CD105-PE, CD274-PE, CD45-APC/Cy7, CD34-PerCP-Cy5.5, CD31-PE and HLA-DR-Pacific Blue (Biolegend, San Diego, USA). Appropriate isotype controls for each of the antibodies were used. Cells cultures a 70–80% confluence were harvested using 0.25% trypsin-EDTA, (Gibco, Life Technologies, Carlsbad, CA, USA) and centrifuged at 1200 rpm × 6 min. Cell pellets were resuspended in 100 µL of 1× PBX with 5% Bovine Serum Albumin (Gibco, Life Technologies, Carlsbad, CA, USA) containing the corresponding antibody. Cell suspension was incubated for 30 min at 4 °C, washed with 1X PBS and resuspended in 200 µL 1× PBS. Flow cytometry analysis were carried out with a FACSCanto II™ instrument (BD, Franklin Lakes, NJ, USA) and data were analyzed with the FlowJo vX.7.0 software package (TreeStar, USA).