摘要
目的对比研究人牙髓干细胞(DPSCs)和根尖乳头干细胞(SCAP)的体外生长特性、增殖及矿化能力。方法采用酶消化法体外培养人DPSCs和SCAP,诱导分化培养基诱导细胞成骨/成牙本质向分化,流式细胞仪检测特异性标记物,茜素红染色检测矿化程度,逆转录PCR(RT-PCR)检测分化标记物表达。结果人DPSCs和SCAP为成纤维细胞样贴壁生长,SCAP增殖率较高;两种细胞表达特异性间充质干细胞(MSCs)标记物:CD34、CD45(-),CD90、CD105、CD146(+),基质细胞抗原1(STRO-1)、八聚体转录因子4(OCT-4)(+),SCAP特异性标记物CD24(+)。成骨诱导3周可形成明显钙化结构,SCAP钙化能力较强。成骨诱导2周2种细胞均可表达分化标记物:骨涎蛋白、骨钙素、牙本质涎磷蛋白,且随诱导时间表达逐渐增加。结论人DPSCs和SCAP均具有间充质干细胞典型特征,可分化为成牙本质样细胞,是牙源性组织工程的可靠干细胞来源。
Objective To compare the growth characteristics,proliferation and osteo/odontogenic differentiation capability of stem cells from human dental pulp（dental pulp stem cells,DPSCs）and apical papilla（stem cells from apical papilla,SCAP）in vitro.Methods Human dental pulp and apical papilla tissues were separated from impacted third molars of young healthy donors at the age of root development and digested by collagenase type Ⅰ and dispase type Ⅱ to derive primitive DPSCs and SCAP.Cells were then induced for osteo/odontongenic differentiation by medium containingβ-glycerophosphate,dexamethasone and KH2PO4.Flow cytometry was utilized to test the expression of specific markers of stem cells,including CD24,CD34,CD45,CD90,CD105,CD146,STRO-1and OCT-4.AR-S was used to display the mineralization structure and RT-PCR was applied to analyze the expression of bone sialoprotein（BSP）,osteocalcin（OCN）and dentine sialophosphoprotein（DSPP）.Results Both DPSCs and SCAP were positive for CD90,CD105,CD146,STRO-1and OCT-4,in percentages varying according to cell type,without expression of CD34 or CD45.Only SCAP expressed CD24 positively.Both cells formed organized mineralization structure after 2 weeks of induction in time-dependent manner,with more mineralization by SCAP and expressed differentiation markers,including BSP,OCN and DSPP.Conclusion Human DPSCs and SCAP possess the characteristics of MSCs and could be differentiated into odontonblast-like cells in vitro.Both cells are approachable stem cell sources for dental tissue engineering.
出处
《重庆医学》
CAS
北大核心
2017年第5期586-589,共4页
Chongqing medicine
基金
国家自然科学基金资助项目(81360161)
云南省教育厅基金资助项目(2015Y153)
关键词
牙髓
干细胞
牙根尖
牙再矿化
组织工程
dental pulp
stem cells
tooth apex
tooth remineralization
tissue engineering
