大白鼠骨髓间质干细胞体外培养体系的建立及分化研究  

Isolation,culture and calcified deposition in vitro of bone marrow mesenchymal stem cells

作　　者：舒胜文[1] 李盛琳[1] 马绪臣[1] 傅嘉[1] 章魁华[1] 

机构地区：[1]北京大学口腔医学院,100081

出　　处：《现代口腔医学杂志》2002年第2期97-100,I001,共5页Journal of Modern Stomatology

基　　金：国家自然科学基金资助项目 (编号 :39870 85 2 )

摘　　要：目的　建立大鼠骨髓间质干细胞 (bonemarrowmesenchymalstemcells ,BMMSCs)体外培养体系。对大白鼠BMMSCs体外培养的条件、供体大白鼠鼠龄等进行了研究。对体外培养的大白鼠BMMSCs的形态学、增殖动力学、成骨的表面标记及体外钙化能力进行了系列研究。方法　取 2月龄SD大鼠胫骨和股骨骨髓进行体外培养。取第二代BMMSCs,绘制生长曲线并计算群体倍增时间。对经成骨诱导剂诱导的BMMSCs的ALP表达及体外钙沉积进行了研究。结果　细胞呈长梭形或多角形 ,细胞表面有不规则的细胞突起。透射电镜可见胞浆有大量的粗面内质网及线粒体 ,细胞表面可见突起。细胞生长曲线呈S形 ,群体倍增时间为 72小时。经诱导分化的BMMSCs可见碱性磷酸酶的表达。vonKossa染色可见散在深染的钙沉积斑块。结论　来源于年轻大鼠股骨骨髓的间质干细胞经体外培养并经诱导后可分化为具有部分成骨特性的细胞 ,可以为体内骨组织工程提供组织细胞。Objective To seek the best way of isolating and culturing bone marrow mesenchymal stem cells (BMMSCs).Histomorphology,ultrastructure,kinetics,expression of ALP and,calcified deposition in vitro were also studied.Methods Bone marrow was collected from femur and tibia of 2 month old male SD rats and seeded in cell culture flask.Their morphology and ultrastructure were analyzed by phase contrast microscope and transmission electron microscope (TEM).Alkaline phosphatase histochemical staining was performed with Gomarri method.Calcified deposition was checked with modified VonKossa method.Results BMMSCs had spindle shape,with irregular processes on cell surface.Large number of rough surfaced endoplasmic reticuli and mitochondria were observed in cells through TEM.Cell doubling time was 72 hours.Induced by culture medium containing dexamethasone,β-glycerphosphate sodium and ascorbic acid,BMMSCs could express ALP obviously.Calcified deposition,which was one characteristics of osteoblast,could also be seen in vitro.Conclusion BMMSCs culture system is established.Through induction,they can differntiate to cells with osteogenesis capability,and may be used to provide cells for bone tissue engineering.

关 键 词：骨髓间质干细胞 细胞培养 体外钙沉积 骨组织工程 

分 类 号：R783.1[医药卫生—口腔医学]