Mouse pre-osteoblasts, MC3T3-E1 - subclone 4 (ATCC CRL-2593), were maintained in αMEM medium supplemented with 10% Fetal Bovine Serum (FBS) at 37 °C and 5% CO2 [19]. To induce osteogenic differentiation, we treated pre-osteoblasts (MC3T3-E1, subclone 4) with ß-Glycerophosphate (10 mM) + Dexamethasone (0.03 g/mL) + Ascorbic Acid (50 μg/mL) (Sigma-Aldrich) culture with 10% FBS [20, 21].
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