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目的通过比较皮质骨和其他不同来源兔成骨细胞的体外培养和部分成骨功能,鉴定并探讨不同来源成骨细胞的增殖、成骨能力及其培养方法。方法取材于3个月龄新西兰大白兔,分别用组织块法和酶消化法分离培养尺骨皮质、髂骨、胫骨骨膜来源的细胞,取髂骨骨髓用密度梯度离心法分离提取干细胞,将细胞分为皮质骨组、骨膜组、松质骨组、干细胞诱导组和干细胞常规组。取第3代细胞进行实验,倒置相差显微镜下观察和结晶紫染色比较细胞形态、MTT法测定细胞增殖曲线、对硝基酚法测碱性磷酸酶(ALP)活性、酶免法测定骨钙素(BGP)活性等。结果普通显微镜下不同来源细胞在形态学上无显著差异;增殖和成骨能力:骨膜组和干细胞常规组增殖最快,皮质骨组和松质骨组居中,干细胞诱导组增殖最慢;干细胞常规组无明显ALP、BGP表达,不同来源细胞的ALP、BGP表达情况大致呈如下关系:骨膜组>皮质骨组>松质骨组>干细胞诱导组。结论皮质骨、骨膜、松质骨、骨髓干细胞均可以培养出成骨细胞,皮质骨来源的成骨细胞成分相对单纯,生命力较旺盛;骨膜细胞在增殖和成骨能力方面有一定优势,但细胞成分相对不纯。
OBJECTIVE: To compare the proliferation and osteogenesis ability of osteoblasts derived from different cortical and other osteoblasts in vitro and in part by their osteogenic function. METHODS: The New Zealand white rabbits (3 months old) were divided into three groups: New Zealand white rabbits (3 months old) were divided into two groups. The osteoblasts, ilium and tibial periosteum cells were isolated and cultured by tissue block method and enzyme digestion method respectively. Bone marrow of iliac bone was separated by density gradient centrifugation. Cortical bone group, periosteum group, cancellous bone group, stem cell induction group and conventional group of stem cells. The third generation cells were used for the experiment. The morphological changes of cells were observed under inverted phase contrast microscope and crystal violet staining. The cell proliferation curve was measured by MTT assay. The alkaline phosphatase (ALP) activity was measured by nitrophenol method. (BGP) activity and so on. Results There was no significant difference in morphology between cells of different origins under ordinary microscope. Proliferation and osteogenic ability were the highest in the periosteum group and the stem cell group, the lowest in the cortical bone group and cancellous bone group, the slowest in the stem cell group, There was no obvious expression of ALP and BGP, ALP and BGP expressions in different cells were as follows: periosteum> cortical bone> cancellous bone> stem cell induction group. Conclusion Cortical bone can be cultured in cortical bone, periosteum, cancellous bone and bone marrow stem cells. The components of cortical bone derived from cortical bone are relatively simple and have a strong vitality. Periosteum has certain advantages in proliferation and osteogenesis, but cells Relatively impure ingredients.