Fitc mouse anti rat cd44h

Expression of surface markers was assessed by flow cytometry using the following conjugated monoclonal antibodies: FITC hamster anti-rat CD29, FITC mouse anti-rat CD44H, FITC mouse anti-rat CD45, FITC mouse anti-rat CD90 (BD Biosciences, San José, CA). MSCs at passage 3 were suspended in PBS and incubated with each antibody at a concentration of 0.5 μg/mL, with unstained MSCs and isotype-control as control.

Passage 3 rat MSCs were digested by 0.25% trypsin, washed twice with PBS, and then resuspended in PBS as single cell suspension. The cells were incubated with fluorescence-coupled antibodies and isotype control antibodies, respectively (0.4 μg antibody: 1 × 10⁵ cells). The antibodies involved are as follows: PE anti-mouse/rat CD29 antibody (BioLegend, USA, catalog number: 102207), FITC anti-rat CD90 antibody (BioLegend, USA, catalog number: 206105), FITC mouse anti-rat CD44H (BD Biosciences, USA, catalog number: 550974), CD34 antibody-FITC (Santa Cruz Biotechnology, USA, catalog number: sc-7324 FITC), CD45 antibody-FITC (Santa Cruz Biotechnology, USA, catalog number: sc-1178 FITC), PE Armenian hamster IgG isotype Ctrl antibody (BioLegend, USA, catalog number: 400907), and FITC mouse IgG1, κ isotype Ctrl antibody (BioLegend, USA, catalog number: 400107). After 30 min of incubation at 4°C in the dark, rat MSCs were washed twice with PBS and resuspended in 300 μl PBS for analysis. Cell fluorescence was examined by a FACScan flow cytometer running CellQuest software.