牙髓干细胞向成牙本质样细胞分化过程中基因表达研究

目的：分离培养来源于人成体牙髓组织的干细胞(dental pulp stem cells，DPSC)，分析研究其在未分化状态及在矿化诱导液中基因表型的变化：方法：采用胶原酶消化法获得人牙髓组织的单细胞悬液，14 d后挑取细胞克隆，分别在普通培养基和矿化诱导培养基中进行培养，利用RT-PCR，图像分析检测细胞基因表型变化。结果。未诱导的DPSC不表达牙本质特异性蛋白(dentin phosphprotein,DSPP)及骨涎蛋白(Bone sialoprotein，BSP)，表达部分成骨(osteocalcin,OCN alkaline phsphatase,ALP)相关表型，低表达转录因子核心结合因子(Cbfa-1)；与末诱导的DPSCs相比较，在成骨诱导中DPSCs表达DSPP和BSP，骨相关基因表达也相应上调。结论：DPsCs在矿化诱导液的作用下向成牙本质样细胞分化，其过程有可能是通过谱系特异表犁的表达或表达上调实现的。

Gene expression in dental pulp stem cell differentiated to odontoblast-like cell

AIM: To isolate and culture stem cells from the human adult dental pulp,then analyze the immunophenotyte of them. METHODS: Mono-cell suspension was separated from the human adult dental pulp with collagenase type I digestion.Cell clones were chosen after 14 days then subcultured in the common culture medium and mineralization induction culture medium.The immunophenotyte of the cells were analyzed with method of RT-PCR and image analysis. RESULTS: DSPP and BSP were not expressed in the uninduced-DPSC.Osteocalcin was partialy expressed and Cbfa-1 was low expressed in the uninduced-DPSC.DSPP and BSP were expressed in the osteocalcin-induced DPSCs.Bone relevant gene expression was increased. CONCLUSION: DPSCs are differentiated to odontoblast by the action of mineralization induction fluid.The process may be completed by the express or express increase of pedigree special phenotype.