Journal: Tissue Engineering. Part A

Article Title: Stem/Progenitor Cell-Mediated De Novo Regeneration of Dental Pulp with Newly Deposited Continuous Layer of Dentin in an In Vivo Model

doi: 10.1089/ten.tea.2009.0518

Figure Lengend Snippet: Multiple lineage differentiation properties of SCAP and DPSCs compared with BMMSCs. (A, D, G) Odonto/osteogenic induction and Alizarin Red staining of matrix mineralization. Insets: uninduced control. All scale bars including insets: 200 μm. (B, E, H) Adipogenic induction and Oil Red O staining of the accumulated lipid droplets in cells. Inset images: higher magnification views of the adipocyte-like cells from the same culture shown in the main images. Scale bars: main images, 100 μm; insets, 50 μm. (C, F, I) Neurogenic induction. βIII-tubulin: red fluorescence; 4',6-diamidino-2-phenylindole dihydrochloride (DAPI): blue fluorescence. Scale bars: (C) 20 μm; (F, I) 30 μm. (J) Myogenic induction and RT-polymerase chain reaction analysis of gene expression. M, RNA from cultured human myoblasts (obtained from Dr. Wesley M. Jackson, NIH/NIAMS, Bethesda, MD); C, uninduced control; m, induction in myogenic medium. SCAP, stem cells from apical papilla; DPSC, dental pulp stem cells; BMMSCs, bone marrow mesenchymal stem cells.

Article Snippet: After fixation in 100% ice-cold methanol, cells were incubated in blocking buffer (32.5 mM NaCl, 3.3 mM Na 2 HPO 4 , 0.76 mM KH 2 PO 4 , 1.9 mM NaN 3 , 0.1% [w/v] bovine serum albumin, 0.2% [v/v] Triton-X 100, 0.05% [v/v] Tween 20, and 5% goat serum) for 30 min followed by the addition of monoclonal mouse anti-human βIII-tubulin antibodies (Promega, Madison, WI) for 1 h at room temperature.

Techniques: Staining, Control, Fluorescence, Polymerase Chain Reaction, Gene Expression, Cell Culture