The Role of Nephronectin on Proliferation and Differentiation in Human Dental Pulp Stem Cells

Figure 8

Evaluation of mineralization capacity in hDPSCs. (a) The mineralization capacity of hDPSCs was confirmed. hDPSCs (passage number 1) were seeded in 6-well plates (tissue culture-treated polystyrene, Iwaki) at the concentration of 3 × 10⁴/well in DMEM supplemented with 10% FBS and penicillin/streptomycin (50 U/mL penicillin; 50 μg/mL streptomycin). Media were changed into DMEM supplemented with 5% FBS, penicillin/streptomycin and mineralization reagent (10 mM β-GP and 50 μg/mL AA) (IM), or PBS vehicle (CM) at day four. The photo shows the alizarin red staining of cells at 23 days. (b) Alizarin red staining of hDPSCs (passage number 4) cultured on various protein-coated nontissue culture-treated polystyrene at day 30. hDPSCs were seeded to modified surfaces at the concentration of 4 × 10³/well (upper) or 6 × 10³/well (lower) in a 24-well plate (nontissue culture-treated polystyrene, Iwaki). Cells were cultured in DMEM supplemented with 10% FBS and penicillin/streptomycin. Mineralization reagent (β-GP and AA) was added at day 4 with 5% FBS containing DMEM. (c) Quantification of alizarin red staining for the lower panel in (b). Significant higher level of mineralization of hDPSCs was detected in Npnt-coated and FCOL1-coated substrate(s). Different symbols represent significant differences, by post hoc Tukey’s HSD test.

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