KMID : 0848120120370040167
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International Journal of Oral Biology 2012 Volume.37 No. 4 p.167 ~ p.173
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A Trial of Screening of Genes Involved in Odontoblasts Differentiation from Human Dental Pulp Stem Cells
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Kim Hyun-Jin
Park Yoon-Kyu
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Abstract
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This study investigated the genes involved in the dif-ferentiation of odontoblasts derived from human dental pulp stem cells (hDPSCs). hDPSCs isolated from human tooth pulp were validated by fluorescence activated cell sor-ting (FACS). After odontogenic induction, hDPSCs were analyzed investigated by Alizaline red-S staining, ALP assay, ALP staining and RT-PCR. Differential display-poly-me rase chain reac tion (DD-PCR) was pe rformed to s c re en differentially expressed genes involved in the differentia-tion of hDPSCs. By FACS analysis, the stem cell markers CD24 and CD44 were found to be highly expressed in hDPSCs. When hDPSCs were treated with agents such as ¥â- glycerophosphate (¥â-GP) and ascorbic acid (AA), nodule formation was exhibited within six weeks. The ALP activity of hDPSCs was found to elevate over time, with a detectable up-regulation at 14 days after odontogenic induc-tion. RT-PCR analysis revealed that dentin sialophospho-protein (DSPP) and osteocalcin (OC) expression had inc-reased in a time-dependent manner in the induction culture. Through the use of DD-PCR, several genes were diffe-rentially detected following the odontogenic induction. These results suggest that these genes may possibly be linked to a variety of cellular process during odontogenesis. Further-more, the characterization of these regulated genes during odontogenic induction will likely provide valuable new in-sights into the functions of odontoblasts.
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KEYWORD
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human dental pulp stem cells, DD-PCR, odon-toblast
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