摘要
目的:研究犬骨髓基质细胞(bone marrow stromal cells,BMSCs)在体外诱导分化为成骨细胞的方法和活性检测。方法:抽取犬骨髓3 mL,使用成骨条件培养液,贴壁法体外培养,将培养的第2、3代细胞进行透射电镜、流式细胞仪、生长曲线、碱性磷酸酶、Von Kossa和免疫组化等检测。结果:第2代细胞碱性磷酸酶染色呈阳性表达;第3代细胞Von Kossa染色、骨桥素、骨涎蛋白和骨钙蛋白呈阳性表达;第3代细胞透射电镜显示高尔基体和线粒体丰富;流式细胞仪检测显示第3代处于S期的细胞比例较第2代增加。结论:犬骨髓基质细胞在体外能定向诱导分化为成骨细胞。
Objective： To study the osteogenic methods and abilities of the dog＇s bone marrow stromal cells （BMSCs） in vitro to estimate its feasibility of being seed cells of bone tissue engineering. Methods： Under general anesthesia, 3 mL bone marrow was aspirated from dog caput femoris. The cells were cultured in delbecco＇s modified eagle medium（DMEM） at 5×10^5 cells/cm^2 and incubated under standard cell culture conditions. The 2nd and 3rd generated cells were collected for studies including transmission electron microscopy（TEM）,fluorescent-activated cell sorter（FACS）,growth curve with MTT method,alkaline phosphatase（ALP） staining.Von Kossa and immunohistologic examination. Result： The expression of ALP of the 2nd generated cell were detected. The 3rd generated cells showed abundant endoplasmic reticulum and mitochondria by TEM, and had more S-staged cells than the 2nd in FACS. The expression of calcic node,osteonctin（ON）, bone sialoprotein（BSP） and osteocalsin（OCN） were proved by Von Kossa staining and immunohistochemical examination. Conclusion： BMSCs have the potentials of being induced into the osteoblasts when cultured in vitro.
出处
《口腔颌面外科杂志》
CAS
2006年第3期209-214,共6页
Journal of Oral and Maxillofacial Surgery
基金
上海市重点学科(优势学科)建设项目(Y0203)
上海市科委项目(03ZR14080
055407034)
国家863计划子课题(2002AA205011)
关键词
骨髓基质细胞
成骨细胞
组织工程
种子细胞
bone marrow stromal cells
osteoblast
tissue engineering
seed cells
