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目的:研究小鼠诱导性多能干细胞(induced pluripotent stem cells,i PSCs)向成骨细胞分化的能力,并观察Osterix对i PSCs成骨分化能力的影响。方法:利用体外细胞培养和病毒转染的方法,结合定量RT-PCR以及组织化学的方法检测在成骨诱导的条件下,i PSCs中成骨相关基因、蛋白的表达变化。结果:小鼠i PSCs经悬滴培养可以形成拟胚体,在成骨诱导条件下可以向成骨细胞分化。病毒转染并不影响i PSCs的多向分化潜能;与对照组相比,转染Osterix的i PSCs形成的矿化结节、碱性磷酸酶活性、成骨相关基因的表达均有明显增加(P<0.05)。结论:在成骨诱导液培养条件下,小鼠i PSCs可以作为种子细胞向成骨细胞分化,并且过表达转录因子Osterix可以进一步促进i PSCs的成骨分化。
Objective: To investigate the ability of induced pluripotent stem cells (iPSCs) to differentiate into osteoblasts and to observe the effect of Osterix on the osteogenic differentiation of iPSCs. Methods: The changes of osteoblast-related genes and proteins in iPSCs under osteogenic conditions were detected by in vitro cell culture and virus transfection combined with quantitative RT-PCR and histochemical methods. Results: Mouse i PSCs can form embryoid bodies by hanging drop culture and differentiate into osteoblasts under osteogenic conditions. Viral transfection did not affect the multidirectional differentiation potential of i PSCs. Compared with the control group, the expression of mineralized nodules, alkaline phosphatase and osteogenesis-related genes were significantly increased in iPSCs transfected with Osterix P <0.05). CONCLUSION: Mouse i PSCs can differentiate into osteoblasts under osteogenic induction fluid culture conditions. Overexpression of Osterix can further promote the osteogenic differentiation of i PSCs.