Mesenchymal Stromal Cell Isolation and Characterization

Mouse mesenchymal stromal cells (mMSC) were isolated, cultured and characterized as previously reported [14 (link)]. Cells were kindly provided by Dr. Osnat Almogi-Hazan, Hebrew University. Bovine mesenchymal stromal cells (bMSC), as we have previously reported [4 (link)], were isolated, cultured and characterized based on generally accepted criteria [3 (link), 15 (link)]. For some experiments and microscopic imaging, cells were preincubated with carboxyfluorescein succinimidyl ester (CFSE; Celltrace, Invitrogen). Briefly, culture medium was removed and cells were treated with 10 μM CFSE in Dulbecco’s Phosphate Buffered Saline (DPBS, Biological Industries, Beit Haemek, Israel) and incubated for 20 min at 37°, 5% CO2. Cells were washed 3 times with a culture medium containing 10% FBS (Gibco), and incubated for further 10 min before harvesting or further processing.