Percp cy5.5 anti human cd 105

The hWJ-MSCs that were used were obtained from passage 3 at 80% confluence. The cells were detached from cell culture flasks (Falcon^®, Coring, Inc., New York, NY, USA) using trypsin/ethylenediaminetetraacetic acid (EDTA) solution 0.05% (Gibco^TM Thermo Fisher Inc., Waltham, MA, USA) and washed with PBS. Then, the hWJ-MSCs were incubated for 30 min at room temperature with saturating concentrations of the following monoclonal antibodies labeled with fluorochromes: PerCP/Cy5.5 anti-human CD 105, FITC anti-human CD90, APC anti-human CD 73, PE anti-human CD 34, PE anti-human CD 45, PE anti-human CD19, PE anti-human CD 11b, PE/Cy7 anti-human HLA-ABC, and PE anti-human HLA-DR (all purchased from BD Biosciences (San Jose, CA, USA). After removing the excess antibody through 3 washes with PBS, the hWJ-MSCs were ana-lyzed according to standard protocols using a flow cytometer (FACS Calibur™, BD Biosciences, San Jose, CA, USA) and FlowJo software (Becton, Dickinson & Company, Franklin, NJ, USA). The cell population was identified, and whether cells were positive or negative for each marker was determined based on the criteria of the International Society for Cell and Genetic Therapy (ISCT).

In this experimental study, MSCs serum-free medium
was ordered from Shanghai Pumao Biotechnology
(Shanghai, China), and trypsin and Dulbecco’s phosphate
buffered saline (DPBS) were ordered from Gibco (Grand
Island, NY, USA). The antibodies of APC-anti-human
CD73, FITC-anti-human CD90, PerCP-Cy5.5- antihuman CD105, PE-anti-human CD34, PE-anti-human
CD45, and PE-anti-human HLA-DR were purchased
from BD Pharmingen (NJ, USA). The CD63 and β-actin
antibodies were ordered from Absin Bioscience (Shanghai,
China) and Cell Signaling Technology (Danvers, MA,
USA), respectively.