联合培养时诱导的内皮细胞对自体骨髓基质干细胞成骨作用的影响  

Osteogenic effects of induced autologous endothelial cells on bone marrow stromal cells of rabbits in co-culture condition

作　　者：孙源[1] 林红[1] 吴子征[2] 陈瑜[1] 董健[1] 

机构地区：[1]复旦大学附属中山医院骨科,上海200032 [2]上海交通大学附属上海市第一人民医院宝山分院骨科

出　　处：《中华创伤骨科杂志》2006年第10期919-923,共5页Chinese Journal of Orthopaedic Trauma

基　　金：国家自然科学基金(30271262);教育部留学回国人员基金(2003-406)

摘　　要：目的探讨由骨髓基质干细胞(BMSCs)诱导的内皮细胞(EC)与自体BMSCs共培养时对BMSCs成骨作用的影响。方法采用密度梯度离心法分离兔BMSCs,培养细胞分为四组：A组(BMSCs组)、B组(BMSCs成骨诱导组)、C组(EC诱导组)及D组(BMSCs和诱导的EC联合培养组),通过干细胞形态、免疫荧光、细胞增殖、碱性磷酸酶活性及骨钙素含量从酶学、组织学及生化等不同方面观察诱导的EC对BM- SCs成骨活性及牛长情况的影响。结果细胞免疫荧光染色证实C组培养诱导的细胞为EC。倒置相差显微镜、HE染色均显示EC与BMSCs混合生长良好。MTF检测结果：各组细胞增殖差异无显著性意义(P＞0．05)。碱性磷酸酶活性和骨钙素含量检测结果：D组明显高于其它各组,差异有显著性意义(P＜0．05)。结论由BMSCs诱导的EC能够增强BMSCs的成骨活性,提高BMSCs的增殖能力。Objective To study the osteogenic effects of induced endothelial cell （EC） on bone marrow stromal cells （BMSCs） of rabbits in co-culture condition. Methods BMSCs were obtained from rabbits by density gradient centrifugation. The adhesive cells were preserved to passage in culture. The cultured cells were divided into four groups： group A （BMSCs）, group B （BMSCs osteogenic induction）, group C （EC induction） and group D （co-cuhure of induced BMSCs and EC） . The cell morphology, immunofluorescence, cell proliferation, alkaline phosphatase （ALP） activity, osteocalcin synthesis were observed to determine the effects of induced autologous EC on the osteogenic potential and cellular compatibility of BMSCs. Results The immunochemical staining showed that the BMSCs were induced into EC in group C. The cellular compatibility of BMSCs and EC was good. The ALP activity and osteocalcin content were obviously higher in group D than in any other groups （ P 〈 0. 05） . The cell proliferation difference was not obvious between groups （ P 〉 0.05）. Conclusions The cellular compatibility of induced osteoblasts and induced EC is perfect. The ECs can significantly increase the viability and ALP activity of induced osteoblasts.

关 键 词：组织工程 骨髓基质干细胞 成骨细胞 内皮细胞 联合培养 

分 类 号：R318.0[医药卫生—生物医学工程]