GSM5114986: KO3_Osteo; Mus musculus; RNA-Seq
Primary MSCs were isolated by flushing the bone marrow of femurs, tibiae and humeri. The bone marrow suspension was cultured in 21cm2 petri dish in alpha minimum Eagle's medium (α-MEM, HyClone) with 10% fetal bovine serum (FBS, Gibco), 100 units/mL penicillin and 100 μg/mL streptomycin. Incubated at 37°C with 5% CO2, the culture medium was changed every 48 hours. MSCs at passage 3 were applied for the subsequent research.  For the osteogenic induction, MSCs were cultured with osteogenic medium supplemented with 50μM ascorbic acid (Sigma), 10nM dexamethasone (Sigma), and 10mM β-glycerophosphate (Sigma). For adipogenic induction, MSCs were treated with adipogenic medium containing 10μg/ml insulin (Sigma), and 1μM dexamethasone (Sigma), 0.5μM isobutylmethylxanthin (Sigma). RNA libiaries were prepared for sequencing using standard Illumina protocols.