目的:比较牙髓、牙周膜和脐带三种不同来源间充质干细胞的(Mesenchymal stem cells,MSCs)三系分化潜能。方法:培养分离人脐带间充质干细胞(Umbilical cord mesenchymal stem cells,UCMSCs)、牙髓干细胞(Dental pulp stem cells,DPSCs)和牙周膜干细胞(Per iodontal ligament stem cells,PDLSCs);倒置显微镜观察细胞形态,体外诱导这3种MSCs向成骨、成脂和成软骨方向分化,通过茜素红染色、油红O染色、阿尔辛蓝染色分别鉴定细胞成骨、成脂和成软骨分化能力,进一步实时荧光定量PCR检测比较3种细胞成骨相关基因OCN和RUNX2及成脂相关基因PPAR-γ的表达水平。结果:这3种MSCs细胞形态略有不同,三者均可向成骨、成脂和成软骨方向分化,但其分化潜能有所差异,DPSCs成骨和成脂向分化能力强,UCMSCs则更适于成脂和成软骨向分化,而PDLSCs成软骨向分化较具优势。结论:DPSCs,PDLSCs和UCMSCs三系分化潜能不同,本研究为干细胞临床转化中选择理想细胞来源提供实验依据。
Objective: To isolate and identify human mesenchymal stem cells from umbilical cord, dental pulp and periodontal ligament(h UCMSC, h DPSC and h PDLSC) and to compare the differentiation potentials of MSCs from the three sources, in order to provide further evidence for MSCs application. Methods: The three type of MSCs were isolated by using explant or enzymatic methods, compared for cell morphology and tri-lineage differentiation potential by using inverted microscope, specifically staining and real-time polymerase chain reaction(RT-PCR), respectively. Results: Cell morphology of h UCMSCs, h DPSCs and h PDLSCs were slightly different, compared with h UCMSCs, h DPSCs and h PDLSCs showed more elongated morphology. The three cell types were differentiated into osteocytes, adipocytes and chondrocytes, but with a difference in differentiation potentials that the osteogenic and adipogenic differentiation potentials of h DPSC were significantly enhanced compared to h UCMSC and h PDLSCs; In contrast, h UCMSC was more able to differentiate into adipogenic and chondrogenic lineage; and h PDLSCs appeared was less able to differentiate into osteogenic and adipogenic lineage, although h PDLSCs have an advantage in chondrogenic differentiation. Conclusions: MSCs from dental pulp, periodontal ligament and umbilical cord possess different differentiation potentials. This study provides an important reference for choice ideal MSCs type in clinical application.