Dual-Lineage Differentiation of Murine BMSC

Primary bone marrow cells were harvested from the tibiae and femurs of CT and SBA mice and grown for 7-10 days in αMEM media (PAN BIOTECH; P04-21050) with 15% fetal bovine serum (PAN BIOTECH; P30-3306), 1% penicillin/streptomycin (PAN BIOTECH; P04-85100) and 1% stable glutamine (PAN BIOTECH; P06-07100). After reaching full confluency, the plated BMSC population was differentiated into both osteoblasts and adipocytes according to the co-differentiation protocol described in a previously published article (30 (link)). The media containing co-differentiation inducers was replaced every two days during the 14 days of the co-differentiation experiment. This culture duration was chosen because of its brevity, which allowed for minimizing the time for cells to change after leaving the in vivo state.