Odontoblast-Like Cells Differentiated from Dental Pulp Stem Cells Retain Their Phenotype after Subcultivation

Figure 3

Immunofluorescence analysis of DSP and DMP-1 localization. Staining of respective positive controls for DSP (placenta) and DMP-1 (mammary gland). In the tooth, a layer of columnar odontoblasts (arrow) surrounds the inner surface of dentin and shows positive staining for both DSP and DMP-1; negative staining in the predentin area was observed (double arrow) and positive staining on the dentin mineralization front (arrowhead). To obtain a detailed protein marker location in cell cultures, they were processed for Alexa 594 immunofluorescence with antibodies to DSP or DMP-1 and Hoechst to stain nuclei after 7, 14, and 21 days of differentiation period (OLC) and 7, 14, and 21 days after reseeding process in fresh cells (EXP-21) and thawed EXP-21 cells (Thawed). Note the intense nuclear staining of DMP-1 and the absence of staining in mesenchymal cells. Bar corresponds to 100 μm.