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. 2019 Mar 18;1(1):56–66. doi: 10.1089/bioe.2018.0005

FIG. 3.

FIG. 3.

Hexokinase treatment rescues OS differentiation in depolarized hMSCs. hMSCs were treated with hex (5 U/mL) to deplete the medium of ATP during OS differentiation. (A) IBSP gene expression was quantified by qPCR after 7 days of hMSC culture in OS medium with 40 mM K+ (HK) and/or hex. Data points are mean normalized expression ± standard deviation, n = 3. (B) Total calcium content was quantified after 21 days of hMSC culture under the same conditions. Data points represent mean calcium concentration (μg/μL) ± standard deviation, n = 6. Different letters over bar graphs represent statistically different groups as determined by one-way ANOVA and the Tukey–Kramer post hoc test, p < 0.05. (C–F) Representative images of cells stained at day 21 for ALP (blue) and calcified mineral (Alizarin Red S). Cells were grown in media supplemented with hex + HK (C), hex (D), HK (E), or in normal OS medium (F). Scale bar = 200 μm. ALP; alkaline phosphatase protein; hex, hexokinase; qPCR, quantitative polymerase chain reaction.