大鼠高产量的破骨细胞分离、诱导培养鉴定与纯化

发布时间：2018-04-02 02:17

本文选题：多核巨细胞　切入点：破骨细胞　出处：《中国医科大学》2006年硕士论文

【摘要】：实验目的正常骨组织,在1mm~3骨中只有2-3个破骨细胞,破骨细胞数量虽然很少但其数量的稳定意义重大,破骨细胞数量的增加使骨吸收大于骨形成从而导致骨质疏松症,破骨细胞数量减少使骨形成大于骨吸收导致骨硬化症,可见破骨细胞在骨骼的形成和骨量的调节方面起着关键作用。目前国内外对破骨细胞研究的切入点主要在于骨质疏松症和骨硬化症两个方面。也有人研究破骨细胞分子生物学的特性和在体外分离培养破骨细胞;分离培养的破骨细胞与成骨细胞混合培养,试图建立模拟体内骨代谢的机制,用以研究骨疾病和抗骨质疏松药物的筛选。还有人已将破骨细胞应用到组织工程骨的评价上,根据培养的破骨细胞与人工制成的工程骨联合培养后形成骨陷凹的差异,即同样的破骨细胞对不同工程骨的噬骨差异来鉴定工程骨质量好坏。有文献报道在成骨细胞和破骨细胞共育体系中,破骨细胞对成骨细胞的功能具有促进作用,其途经是破骨细胞分泌了某些蛋白质,弥散到隔膜的另一侧影响了成骨细胞的功能,说明成骨细胞和破骨细胞之间相互促进相互影响。目前制备的组织工程骨只注重单一成骨细胞种植到骨支架上,而忽略了破骨细胞对成骨细胞的促进影响及其对骨支架的改造朔形作用。相信在不远的将来破骨细胞应用到骨组织工程会产生深远的影响。目前破骨细胞分离培养的方法通常由较大动物禽类(兔和鸡等)四肢长骨机械分离直接获得,对于较小的动物如大鼠由于在体存在的破骨细胞数量较少,所以用经典的方法想获得较多的破骨细胞是不可能的。一般采取诱导成年大鼠骨髓单核细胞形成破骨细胞。各种方法获得的破骨细胞中往往混杂有其他贴附细胞。本研究目的就是要用改进的新技术获得大鼠高产量高纯度破骨细胞,同时探讨破骨细胞分泌的明胶酶参与骨陷凹形成机制,为在体外研究破骨细胞生物特性、骨吸收功能及进一步应用到骨组织工程提供
[Abstract]:Experimental purposeIn normal bone tissue, there are only 2-3 osteoclasts in the 1mm~3 bone. Although the number of osteoclasts is small, the number of osteoclasts is of great significance. The increase of osteoclasts makes bone resorption larger than bone formation, which leads to osteoporosis.The decrease in the number of osteoclasts makes the formation of bone larger than that of bone resorption leading to ossification. Osteoclasts play a key role in the formation of bone and the regulation of bone mass.At present, the entry point of osteoclast research at home and abroad mainly lies in osteoporosis and ossification.There are also studies on the molecular biological characteristics of osteoclasts and the isolation and culture of osteoclasts in vitro, and the co-culture of osteoclasts and osteoblasts in an attempt to establish mechanisms that mimic bone metabolism in vivo.To study bone disease and the screening of anti-osteoporosis drugs.Others have applied osteoclasts to the evaluation of tissue engineered bones, based on the difference between cultured osteoclasts and artificial engineered bone to form a concave bone.That is, the same osteoclast to different engineering bone to identify the quality of bone.In the co-culture system of osteoblasts and osteoclasts, it has been reported that osteoclasts promote the function of osteoblasts by the way that the osteoclasts secrete some proteins and diffuse to the other side of the septum to influence the function of osteoblasts.These results suggest that osteoblasts and osteoclasts interact with each other.At present, the tissue engineered bone only focuses on the single osteoblast implanted on the bone scaffold, but neglects the influence of osteoclasts on osteoblasts and the remodeling of bone scaffolds.It is believed that the application of osteoclasts to bone tissue engineering will have a profound impact in the near future.At present, the methods of osteoclast isolation and culture are usually obtained by mechanical isolation of the long bones of the limbs of larger animal birds (rabbits, chickens, etc.). For smaller animals such as rats, the number of osteoclasts is relatively small due to the presence of osteoclasts in vivo.So it's impossible to get more osteoclasts in the classic way.In general, adult rat bone marrow monocytes are induced to form osteoclasts.The osteoclasts obtained by various methods are often mixed with other adherent cells.The aim of this study was to obtain rat osteoclasts with high yield and high purity by using the improved new technique, and to explore the mechanism of osteoclasts secreting gelatinase in the formation of osteoclasts, so as to study the biological characteristics of osteoclasts in vitro.Bone resorption function and its further application to bone tissue engineering
【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329

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