人骨髓间质干细胞向成骨细胞分化的形态学研究  

Morphological characteristics of human mesenchymal stem cells in the process of osteogenic differentiation

作　　者：庞永刚[1] 崔鹏程[1] 陈文弦[1] 

机构地区：[1]解放军第四军医大学唐都医院耳鼻咽喉科,陕西省西安市710038

出　　处：《中国临床康复》2004年第11期2036-2038,共3页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金(30171007)~~

摘　　要：目的:研究人骨髓间质干细胞(hMSCs)体外培养并向成骨细胞表型转化过程中的形态学改变。方法:在成骨特定诱导培养液中培hMSCs。在相差显微镜和电子显微镜下观察细胞形态,苏木精-伊红染色观察形态,组织化学法检测碱性磷酸酶(ATP)、Ca结节染色,免疫组化检测Ⅰ型胶原(COLⅠ)、骨钙素(OC)表达。结果:诱导培养的hMSCs形态由成纤维样梭形向多边形转变,透射电镜观察见大量扩张的粗面内质网、高尔基体、线粒体和钙盐沉积,扫描电镜见细胞表面有大量钙盐颗粒。诱导培养后7,14,21d可见ALP染色、COLⅠ免疫组化染色阳性,诱导培养后14,21d可见OC免疫组化染色、Ca结节染色阳性。结论:hMSCs在特定培养液诱导下可表现为典型的成骨细胞形态,可作为骨组织工程的种子细胞。AIM:To investigate the in vitro culture of human mesenchymal stem cells(hMSCs) derived form bone marrow and their morphological changes in the process of oste ogenic differentiation. METHODS:hMSCs derived from bone marrow were cultured in the special medium to induce the osteogenic differentiation.Morphological changes of the hMSCs were o bserved under phase contrast microscope and electron microscope.The hMSCs were s tained with hematoxylin and eosin(HE) to observe their morphological characteris tics.Alkaline phosphatase(ALP) and the node of calcium were detected by using hi stochemical method,and the expressions of type-I collegan(COLⅠ) and osteocalci n(OC) were measured with immunohistochemical staining. RESULTS:The hMSCs cultured in the medium changed from the spindle-shape to po lygon-shape.A great number of rough endoplasmic reticula,Golgi apparatus,mitoch ondria,calcium deposition were observed under transmission electron microscope,a nd many calcium granules on the surface of the cells and in the matrix were obse rved under scanning electron microscope.After induction and culture,the positive expressions of ALP and COL Ⅰ were detected on day 7,14 and 21,and the positiv e expressions of OC and the node of calcium were found on day 14 and 21.CONCLUSION:Introduced in the specific culture medium,hMSCs derived from bone m arrow can manifest the typical morphological characteristics of osteoblasts,prov iding seed cells for bone tissue engineering.

关 键 词：骨髓间质干细胞 成骨细胞 细胞分化 形态学 生物医学工程 碱性磷酸酶 检测 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学]