细胞凝集态在钙结节形成过程中的显微结构观察  

Microstructure observation of cellular condensations in calcium nodules

作　　者：潘锋[1,2] 柏树令[1] 

机构地区：[1]中国医科大学基础医学院解剖学教研室 [2]基础医学院生物材料研究室,沈阳110001

出　　处：《解剖学报》2011年第3期399-402,共4页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(81000791);辽宁省教育厅高校资助基金资助项目(L2010580)

摘　　要：目的观察兔骨髓基质细胞来源的细胞凝集态在体外钙结节形成过程的形态变化。方法通过全骨髓培养方法分离培养兔骨髓基质细胞,并在培养瓶中进行扩增,使用成骨诱导液作为培养基;再通过四环素标记、碱性磷酸酶染色、茜素红染色、扫描电镜及透射电镜方法,研究细胞凝集态在钙结节形成过程中的形态变化。结果荧光显微镜下观察可见,四环素标记的钙结节发出金黄色荧光;光镜下见结节中的细胞凝集态与周边铺壁细胞相比呈碱性磷酸酶及茜素红的强阳性反应;扫描电镜下见结节中的凝集态细胞均呈球状生长并分泌基质;透射电镜下见细胞中基质小泡丰富。结论骨髓来源的基质细胞在体外形成的钙结节需要三维空间状态下细胞的高度凝集。Objective To observe the morphological changes of cellular condensations from marrow stromal cells in calcium nodules, and to provide experimental data for further studying bone regeneration. Methods Rabbit marrow stromal cells were isolated through whole marrow culture method and amplified in flasks, using the osteohlast inducing conditional media served as the culture media. Morphological changes of cellular condensations in minerized nodules were investigated by tetracycline labelling, alkaline phosphatase staining （ALP）, alizarin red S staining （ARS）, scanning electron microscope （SEM）and transmission electron microscope （TEM）. Results Calcium nodules with cellular condensations revealed bright golden yellow fluorescence under fluorescence microscope when labelled with tetracycline. Histoehemical study with ARS and ALP staining showed positive calcium reaction and positive alkaline phosphatase reaction compared with pheripheral spreading cells, respectively. Under SEM, condensed cells with spherai shape secreted abundant granules heaped up in the cell body. Rich matrix vesicles could be seen under TEM. Conclusion Calcium formation in vitro from marrow stromal cells needs highly condensed cells in three dimensional space.

关 键 词：细胞凝集态 钙结节 扫描电镜 透射电镜 兔 

分 类 号：R318.12[医药卫生—生物医学工程]