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In Vitro Chondrogenesis Transformation Study of Mouse Dental Pulp Stem Cells

机译:小鼠牙髓干细胞的体外软骨生成转化研究

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摘要

A major challenge in the application of mesenchymal stem cells in cartilage reconstruction is that whether the cells are able to differentiate into fully mature chondrocytes before grafting. The aim of this study was to isolate mouse dental pulp stem cells (DPSC) and differentiate them into chondrocytes. For this investigation, morphological, molecular, and biochemical analyses for differentiated cells were used. To induce the chondrocyte differentiation, DPSC were cultured in chondrogenic medium (Zen-Bio, Inc.). Based on morphological analyses using toluidine blue staining, proteoglycan products appear in DPSC after 21 days of chondrocyte induction. Biochemical analyses in differentiated group showed that alkaline phosphatase activity was significantly increased at day 14 as compared to control (P < 0.05). Cell viability analyses during the differentiation to chondrocytes also showed that these cells were viable during differentiation. However, after the 14th day of differentiation, there was a significant decrease (P < 0.05) in the viability proportion among differentiated cells as compared to the control cells. In RT-PCR molecular analyses, mouse DPSC expressed Cd146 and Cd166 which indicated that these cells belong to mesenchymal stem cells. Coll I and Coll II markers showed high expression after 14 and 21 days, respectively. In conclusion, this study showed that DPSC successfully differentiated into chondrocytes.
机译:间充质干细胞在软骨重建中的应用中的主要挑战是细胞是否能够在移植前分化为完全成熟的软骨细胞。这项研究的目的是分离小鼠牙髓干细胞(DPSC),并将其分化为软骨细胞。为了进行这项研究,使用了分化细胞的形态,分子和生化分析。为了诱导软骨细胞分化,在软骨形成培养基(Zen-Bio,Inc。)中培养DPSC。根据使用甲苯胺蓝染色的形态分析,软骨细胞诱导21天后,蛋白聚糖产物出现在DPSC中。分化组的生化分析表明,与对照组相比,第14天的碱性磷酸酶活性显着增加(P <0.05)。分化为软骨细胞期间的细胞生存力分析还表明,这些细胞在分化过程中是可行的。然而,在分化的第14天之后,与对照细胞相比,分化的细胞之间的生存力比例显着降低(P <0.05)。在RT-PCR分子分析中,小鼠DPSC表达Cd146和Cd166,表明这些细胞属于间充质干细胞。 Coll I和Coll II标记分别在14天和21天后显示高表达。总之,这项研究表明DPSC成功地分化为软骨细胞。

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