Pe conjugated anti mouse cd29

We quantitatively investigated the population difference between WT and GM3^−/− mice. MSCs (1 × 10⁶) were stained with FITC-conjugated anti-mouse CD45 (eBioscience, USA), PE-conjugated anti-mouse CD29 (eBioscience), or PE-conjugated CD86 antibodies (eBioscience). Cell surface marker expression was analyzed by flow cytometry (FACSCanto, BD Biosciences, San Jose, CA, USA).

Matrigel and the surrounding hind limb muscle were harvested from euthanized mice and enzymatically digested with Collagenase A (1 mg/mL; Roche Life Science) and Dispase (2.5 U/mL; BD Biosciences) for 2 h at 37 °C. The retrieved cells were incubated with PerCP-conjugated anti-mouse CD45 (1:100; BD Biosciences), PE-conjugated anti-mouse CD11b (1:100; BD Biosciences), PE-conjugated anti-mouse F4/80 (1:50; AbD Serotec), FITC-conjugated mLy6G (1:50; eBiosciences) and APC-conjugated Ly6C(1:100, eBioscience). Flow cytometric analyses were performed using a Guava easyCyte 6HT/2L flow cytometer (Millipore Corporation, Billerica, MA) and FlowJo software (Tree Star Inc., Ashland, OR). To determine ECFC and MPC retention, the retrieved cells were stained with PerCP-conjugated anti-mouse CD45, FITC-conjugated anti-human CD31 (1:10 BD Biosciences), and PE-conjugated anti-human CD90 (1:100 BD Biosciences). Mouse endothelial and stromal cell recruitment was stained by PerCP-conjugated anti-mouse CD45, PE-conjugated anti-mouse CD29 (1:100 eBioscience) and APC-conjugated anti-mouse CD31 (1:100 eBioscience).