Thiazolidinedione-induced lipid droplet formation during osteogenic differentiation

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   Figure 1

   MSC surface marker expression. (A) SC MSCs: CD90+CD45−. (B) PV MSCs: CD90+CD45−. A total of 15 000 events were recorded, of which 9948±401 and 8977±745 were gated as SC and PV respectively.
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   Figure 2

   Adipogenesis: lipid accumulation. (A) Oil red O staining (day 14). (B) Lipid accumulation under different medium conditions without TZD treatment. (C) TZD treatment in AM with PPARγ antagonist (GW9662). (D) TZD treatment in AM without indomethacin. (E) TZD treatment in AM without indomethacin and with added GW9662. Statistical analysis: ^*P<0.01, ***P<0.001 indicate significant difference from AM (B) ^$P<0.05, ^$$P<0.01, ^$$$P<0.001 indicate significant difference between cell types within the same media and TZD treatment conditions (B and D). Factorial ANOVA with Tukey's post hoc test. ^#P<0.05, ^##P<0.01, and ^###P<0.001 indicate significant difference from control media within the same cell type. ⁺P<0.05 and ⁺⁺⁺P<0.001 indicate significant difference between TZD treatments in the same cell type. ^$P<0.05 and ^$$$P<0.001 indicate significant difference between cell types within the same media and TZD treatment conditions. A full colour version of this figure available via http://dx.doi.org/10.1530/JOE-14-0425.
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   Figure 3

   Osteogenesis: mineralisation. (A) No evidence of mineralisation in SGM with or without PPARγ antagonist (GW9662). (B) Mineralisation in different medium conditions without TZD treatment. (C) Alizarin red S staining (day 21) demonstrating mineralisation with lipid droplets evident with TZD treatment. (D) Mineralisation with TZD treatment in OM. (E) Mineralisation with TZD treatment in OM+GW9662. (F) Lipid accumulation with TZD treatment in OM. (G) Lipid accumulation with TZD treatment in OM+GW9662. Statistical analysis: factorial ANOVA with Tukey's post hoc test. ^##P<0.01 and ^###P<0.001 indicate significant difference between SGM and other medium conditions (B). **P<0.01 and ***P<0.001 indicate significant difference between OM and OM+GW9662 (B) and significant TZD treatment effect (D, E, F and G) in the same cell type. ^$$$P<0.001 indicate significant difference between SC and PV MSCs in the same treatment conditions. A full colour version of this figure available via http://dx.doi.org/10.1530/JOE-14-0425.
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   Figure 4

   Pro-adipogenic gene expression. The relative mRNA expression of Pparγ2 (A and B), Ap2 (C and D), and Adipsin (E and F) in SC MSCs and PV MSCs. Statistical analysis: factorial ANOVA with Tukey's post hoc test. *P<0.05 and **P<0.01 indicate significant treatment effect compared with AM at the same time point. Note: values are expressed as the relative expression of Pparγ2, Ap2, or Adipsin compared with the housekeeping gene, Arbp.
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   Figure 5

   Pro-osteogenic gene expression. The relative mRNA expression of Msx2 (A and B), Collagen I (C and D), and alkaline phosphatase (E and F) in SC MSCs and PV MSCs. Statistical analysis: factorial ANOVA with Tukey's post hoc test. *P<0.05 indicate significant treatment effect compared with OM at the same time point. Note: values are expressed as the relative expression of Msx2, Collagen I, or Alp compared with the housekeeping gene, Arbp.

• © 2014 Society for Endocrinology