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. 2014 Sep 8;34(5):1201–1208. doi: 10.3892/ijmm.2014.1926

Figure 1.

Figure 1

Rat calvarial osteoblast cultures form bone more abundantly in αMEM. All cells were cultured in 50 μg/ml ascorbate and 2 mM β-glycerophosphate. (A) The images are representative, whole well reflective light scans of rat osteoblast cell layers cultured using DMEM or αMEM supplemented with 10% fetal calf serum (FCS) or heat-inactivated FCS (HI FCS), with or without 10 nM dexamethasone. Cell layers are either unstained (white), stained with alizarin red to show bone mineralisation (red) or for tissue non-specific alkaline phosphatase (TNAP) expression (purple). Scale bar, 5 mm. (B) The level of bone mineralisation was 3-fold higher in the cells cultured in αMEM compared to DMEM. In cultures without dexamethasone, bone mineralisation was 85% lower (αMEM) or completely absent (DMEM). Heat inactivation of FCS did not have an effect on the level of bone formation. (C) Levels of TNAP expression were 2.5-fold higher in the cells cultured in αMEM. The absence of dexamethasone had no effect on TNAP expression when the cells were grown in αMEM, but TNAP expression was reduced by 3- to 4-fold in the cells cultured in DMEM. Heat inactivation of FCS did not affect TNAP expression. Values are the means ± SEM (n=6 replicate wells), ***p<0.001.