成人骨髓基质细胞的体外培养和鉴定

发布时间：2018-09-17 06:52

【摘要】：骨髓是一个具有多种细胞成分的复杂器官,有造血及成骨能力。骨髓细胞的成骨能力来源于骨髓基质系统的基质干细胞(Stromalstem cell),在体外,骨髓基质细胞的成骨作用已得到肯定。由于骨髓具有取材方便、对机体损伤小的特点,成为骨组织工程最有应用前景的种子细胞。但骨髓基质细胞具有多分化潜能,且有多种成分,因此,骨髓基质细胞的纯化和对其定向诱导分化为成骨细胞,是其应用于骨组织工程的前提。本实验是在动物实验的基础上,探讨一种通过换液纯化骨髓基质细胞,使用不同浓度地塞米松以寻找一对骨髓基质细胞的增殖和分化均较适宜的浓度,并在适宜浓度的条件培养基条件下使其定向分化,旨在建立一种简便易行的成人骨髓基质细胞体外培养方法,为进一步研究成骨细胞及骨组织工程提供技术平台。目的:建立一种成人骨髓基质细胞在体外培养条件向成骨细胞转化的方法,探讨简易的成人成骨细胞体外培养方法,为进一步研究成骨细胞及骨组织工程提供技术平台。方法:手术中抽取健康成人骨髓组织,采用全骨髓培养法在体外进行培养,细胞汇合后改用含不同浓度地塞米松的培养基培养后观察细胞的增殖和分化情况。再采用适宜浓度地塞米松、β-甘油磷酸钠和维生素C的条件培养基进行传代培养,观察细胞增殖和分化情况,并对细胞进行鉴定。结果:通过对不同浓度地塞米松对骨髓基质细胞增殖与分化的影响观察,认为10~(-8)mol/L的浓度为比较合适的浓度。并采用全骨髓法在体外培养的成人成骨细胞表现成骨细胞的形态特征,细胞ALP阳性
[Abstract]:Bone marrow is a complex organ with multiple cellular components, with hematopoietic and osteogenic capacity. The osteogenic ability of bone marrow cells is derived from stromal stem cell (Stromalstem cell), of bone marrow stromal system in vitro, and the osteogenic effect of bone marrow stromal cells has been confirmed. Bone marrow is the most promising seed cell for bone tissue engineering because of its advantages of easy extraction and less damage to body. However, bone marrow stromal cells have multiple differentiation potential and many components. Therefore, the purification and directional induction of bone marrow stromal cells into osteoblasts is the premise of its application in bone tissue engineering. On the basis of animal experiments, we studied a suitable concentration of dexamethasone for the proliferation and differentiation of bone marrow stromal cells (BMSCs), which were purified by fluid exchange, and used dexamethasone in different concentrations to find out the suitable concentration for the proliferation and differentiation of bone marrow stromal cells. In order to establish a simple culture method of adult bone marrow stromal cells in vitro and provide a technical platform for further study of osteoblasts and bone tissue engineering. Objective: to establish a method of transforming adult bone marrow stromal cells into osteoblasts in vitro, and to explore a simple method of adult osteoblasts culture in vitro, and to provide a technical platform for further research on osteoblasts and bone tissue engineering. Methods: the healthy adult bone marrow tissue was extracted during operation and cultured in vitro by whole bone marrow culture method. The proliferation and differentiation of the cells were observed after the cells were confluent and cultured in different concentrations of dexamethasone medium. Then the cells were subcultured in the suitable concentration of dexamethasone, 尾 -glycerophosphate and vitamin C, the proliferation and differentiation of the cells were observed, and the cells were identified. Results: the effects of different concentrations of dexamethasone on proliferation and differentiation of bone marrow stromal cells were observed. It was concluded that the concentration of 10 ~ (-8) mol/L was the appropriate concentration. The adult osteoblasts cultured in vitro showed the morphological characteristics of osteoblasts and ALP positive cells by whole bone marrow method.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R329.2

【引证文献】