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Osteogenic differentiation

JB Jasmin Bagge
LB Lise Charlotte Berg
JJ Jennifer Janes
JM James N. MacLeod
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Passage 4 cells were seeded as monolayer cultures at a density of 25,000 cells per well in 6-well plates (Thermo Fisher Scientific, Waltham, Ma, USA). Initially, the cells were cultured in expansion medium at 37 °C and 5% CO2 with media change every 2–3 days. Upon ~ 90% confluence, expansion medium was changed to osteogenic induction medium in three of the wells, while the other three wells were kept in expansion medium as a non-induced control. Osteogenic induction medium consisted of DMEM (1 g/L glucose), 10% (v/v) FBS, 1% (v/v) P/S, 100 nM dexamethasone, 0.05 mM ascorbic acid (A2P), and 10 mM β-glycerophosphate disodium salt hydrate (Sigma-Aldrich, St. Louis, MI, USA) as previously described [15, 48, 62]. Medium in both groups was changed every 3 days with 2 mL medium per well.

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