年龄因素对人牙周膜干细胞培养和性能的影响

The effects of aging on the isolation and characteristics of human periodontal ligament stem cells from human impacted third molars

目的:探讨年龄因素对人牙周膜干细胞(periodontal ligament stem cells,PDLSCs)体外培养及其生物学特性的影响。方法:收集临床<30岁、>50岁具有完整牙根的第三磨牙各4个,分别刮取根面残留牙周膜进行PDLSCs分离培养,观察其细胞克隆形成能力、MTT方法检测生长曲线、流式细胞仪分析细胞表面分子,并进行体外成骨、成脂诱导,对比分析PDLSCs多向分化能力。结果:两组第三磨牙中均可培养出具有明显间充质干细胞(mesenchymal stem cell,MSC)特性的PDLSCs,但>50岁组获得的PDLSCs克隆形成率显著降低(P<0.05),其增殖、分化能力亦较<30岁组明显减弱(P<0.05)。结论:年龄因素对PDLSCs体外培养及其生物学特性具有明显影响;PDLSCs的基础和临床研究中,应充分考虑到病人年龄因素。

AIM： To investigate the effects of aging on the isolation and characteristics of human periodontal ligament stem cells（PDLSCs） from human impacted third molars.METHODS： Eight impacted third molars from 8 different donors（without caries or periodontal disease） were collected in clinic and divided into two groups based on the donors＇ ages： A group（donor age less than 30 years） and B group（donor age more than 50 years）.The remaining periodontal ligament tissues were used for cell culture.Primary PDLSCs were obtained and sub-cultured for colony-forming assay.Cell surface markers were determined by flow cytometric analysis,cell proliferation was investigated by MTT method,and finally,after 2 weeks（adipogenic differentiation） or 3 weeks（osteogenic differentiation） of induction,the cells were stained with oil red O stain or alizarin red respectively for osteogenic and adipogenic differentiation assay.The data were contrastively evaluated and quantified for statistical analysis.RESULTS： PDLSCs with mesenchymal stem cell（MSC） properties were successfully obtained from all the 8 teeth.The obtained data showed that the proliferation and differentiation abilities of PDLSCs from B group were relatively low as compared with those from A group（P0.05）. CONCLUSION： Human PDLSCs can be successfully isolated from PDL tissue derived from aged donors,but age-related changes of the MSC properties should be taken into account whenever they are intended to be used for investigations.