Fitc anti human cd105 antibody

The characteristics of hUC-MSCs were evaluated using flow cytometry. Passage 5 hUC-MSCs were trypsinized and centrifuged. Next, 1 × 10⁶ cells were resuspended in 100 μL of cold phosphate-buffered saline (PBS) per sample and then stained with APC anti-human CD166 antibody (Biolegend, 343906), APC anti-human CD29 antibody (Biolegend, 303008), FITC anti-human CD90 (Thy1) antibody (Biolegend, 328108), FITC anti-human HLA-DR antibody (Biolegend,327006), FITC anti-human CD73 (Ecto-5’-nucleotidase) antibody (Biolegend, 344016), FITC anti-human CD105 antibody (Biolegend, 323204), FITC anti-human CD45 antibody (Biolegend, 304006), respectively. After 15 min, the cells were washed and resuspended in 100 μL PBS. The cells were analyzed using a flow cytometer (BD Biosciences, FACSCelesta™). In total, 50,000 events were collected per sample. The data were analyzed using FlowJo software (Tree Star Inc., Ashland, OR, USA).

CAFs were processed to cell suspension and counted. The antibodies were added as manufacturer’s guidance (FITC anti-human CD105 Antibody, Biolegend, USA, Cat: 323203. FITC anti-human CD90 Antibody, Biolegend, USA, Cat: 328107. PE anti-human CD146 Antibody, Biolegend, USA, Cat: 361005. PE anti-human CD34 Antibody, Biolegend, USA, Cat: 378603. PE anti-human HLA-DR Antibody, Biolegend, USA, Cat: 327007. PE anti-human CD73 Antibody, Biolegend, USA, Cat: 344003). After incubated for 30 min at 37℃ in dark conditions, the samples were analyzed or sorted. Then the cells were collected and cultured in DMEM/F12 media supplemented with 10% FBS and 1% penicillin/streptomycin.