Anti human cd105

Cells were treated with 0.05% trypsin-EDTA (Invitrogen) for 3 minutes in a 37 °C incubator. Cells were dissociated by gentle pipetting and filtered through a 40-μm cell strainer. The dissociated cells were immediately resuspended at approximately 2 × 10⁵ cells per ml in PBA (1% bovine serum albumin, 0.02% NaN₃ in PBS) and incubated with each MAbs (anti-human CD14-FITC (BD Biosciences, Cat. No. 557153), anti-human CD31-FITC (BD Biosciences, Cat. No. 560984), anti-human CD105 (BD Biosciences, Cat. No. 561443), anti-human CD117-PE (BD Bioscience, Cat. No. 340529), anti-human CD146-FITC (BD Bioscience, Cat. No. 560846), anti-human VEGFR2-FITC (Miltenyi Biotec, Cat. No. 130-105-303), anti-human CD144-FITC (BD Bioscience, Cat. No. 560874), anti-Human CD34-FITC (BD Bioscience, Cat. No. 555821), anti-human CD45-FITC (Biolegend, Cat. No. 368508) for 30 minutes at 4 °C. After washing twice with PBA, propidium iodide (PI)–negative cells were analyzed for the antibody binding using FACSCalibur (BD Biosciences – Immunocytometry Systems, and Cell Quest software (BD Biosciences - immunocytometry Systems).

Established ASCs from both healthy donors were examined for cellar morphology during passage, and characterized for CD markers characterized with mesenchymal stem cell via flow cytometry. The primary antibodies for analysis were anti-human CD73 (BD Biosciences, San Jose, CA, USA), anti-human CD90 (BD Biosciences, San Jose, CA, USA) and anti-human CD105 (BD Biosciences, San Jose, CA, USA) which were used at the manufacturer's recommendations.