Isolation and culture of MSCs from mouse bone marrow were performed according to our previous protocol.22,24 Briefly, whole bone marrow cells from femora and tibia were seeded, incubated overnight, and rinsed with phosphate-buffered saline (PBS) to remove the nonadherent cells. The adherent cells were cultured with alpha-minimum essential medium supplemented with 20% fetal bovine serum (FBS), 2 mmol·L−1 L-glutamine, 55 μmol·L1 2-mercaptoethanol, 100 U·mL−1 penicillin, and 100 μg·mL−1 streptomycin (all from Invitrogen, USA) at 37 °C in a humidified atmosphere of 5% CO2. MSCs were digested with 0.25% trypsin (Invitrogen, USA) and passaged for functional experiments after seeding at appropriate densities.

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