Isolation and Characterization of Rat BM-MSCs

Isolation of rat BM-MSCs was performed as described previously^{9 (link)}. In brief, after male Sprague Dawley rats (Sankyo Lab Service Corp.) were sacrificed, BM was flushed out from removed tibias and femurs. Marrow was cultured in alpha-minimal essential medium (Thermo Fisher Scientific, Lafayette, CO, USA) with 15% fetal bovine serum (FBS) (CCB, NICHIREI BIOSCIENCE, Tokyo, Japan) and 1% penicillin streptomycin (PS) (Thermo Fisher Scientific). The medium was first changed 24 h after seeding and then twice weekly. BM-MSCs that were harvested after up to three passages and that were positive for CD90 and negative for CD45 and CD11b^{44 (link)} were used for further experiments.