人胚胎来源多潜能干细胞向成骨细胞分化的研究

发布时间：2018-06-02 23:43

本文选题：胚胎多潜能干细胞 + 成骨细胞　；参考：《吉林大学》2010年硕士论文

【摘要】： 由于外伤、感染、肿瘤切除以及发育异常等原因造成的骨组织缺损的修复,一直是骨科领域所面临的重大问题。近年来,随着骨组织工程研究的不断发展,种子细胞的来源、培养、诱导、增殖、分化等研究逐渐成为骨组织工程研究的热点。由于干细胞在特定的环境下,具有能大量增殖、分化为各种组织、器官的潜能性以及表达外源目的基因等优点,并且可以在特定的条件下定向向成骨细胞分化。因此,利用干细胞的多向分化潜能治疗骨组织损伤,有着广阔的研究前景。本实验以人胚胎肝脏组织中分离出的多潜能干细胞(胎肝干细胞)为研究对象,利用维生素C、地塞米松、β-甘油磷酸钠,在体外将其诱导分化为成骨细胞,并通过茜素红染色、碱性磷酸酶染色、钙结节染色、RT-PCR、免疫细胞化学等方法,对诱导后的细胞进行鉴定,探讨胎肝干细胞分化为成骨细胞的潜能及其分化机制。结果表明,体外环境下在适当浓度的维生素C、地塞米松、β-甘油磷酸钠的共同作用下,胎肝干细胞可以分化为成骨细胞,茜素红染色、碱性磷酸酶染色等检测结果呈阳性,RT-PCR检测到成骨细胞OPN基因的表达,免疫细胞化学检测结果也呈阳性,为今后干细胞用于临床治疗骨组织缺损提供一定的理论依据和实践依据。
[Abstract]:The repair of bone defects caused by trauma, infection, tumor excision and abnormal development has always been a major problem in orthopedics. In recent years, with the development of bone tissue engineering, the research on the origin, culture, induction, proliferation and differentiation of seed cells has gradually become a hot spot in bone tissue engineering. Stem cells have the advantages of proliferation, differentiation into various tissues, organ potential and expression of exogenous target genes in a specific environment, and can be directed to osteoblast differentiation under specific conditions. Therefore, the multidirectional differentiation potential of stem cells in the treatment of bone injury has a broad research prospect. In this study, multipotent stem cells (FHSCs) isolated from human embryonic liver tissues were induced to differentiate into osteoblasts by vitamin C, dexamethasone, 尾 -glycerophosphate in vitro and stained with alizarin red. Alkaline phosphatase staining (ALP), calcium nodule staining (RT-PCR) and immunocytochemistry were used to identify the induced cells, and to investigate the potential of fetal liver stem cells to differentiate into osteoblasts and the mechanism of differentiation. The results showed that fetal liver stem cells could differentiate into osteoblasts and stained with alizarin red under the combined action of vitamin C, dexamethasone and 尾 -glycerophosphate in vitro. The expression of OPN gene in osteoblasts was detected by RT-PCR and the results of immunocytochemistry were also positive by alkaline phosphatase staining, which provided some theoretical and practical basis for clinical treatment of bone tissue defects by stem cells.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R329

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