中文摘要：

目的：构建可稳定表达成纤维细胞生长因子18蛋白的真核表达载体，在体外观察成纤维细胞生长因子18对兔骨髓基质干细胞体外增殖的影响。 方法：实验于2002—10／2004—03在西安第四军医大学全军骨肿瘤研究所及口腔医学院完成。①将pGEM—T-成纤维细胞生长因子18-3和pSecTag2B表达载体分别用KpnⅠ和NotⅠ酶切后重组pSecTag2B-成纤维细胞生长因子18质粒。（当脂质体介导下转染哺乳动物细胞COS-7，获得稳定表达后用细胞上清刺激骨髓基质干细胞。③通过噻唑蓝检测细胞增殖情况。 结果：①成功构建pSecTag2B-成纤维细胞生长因子18真核表达载体（电泳可见540bp的特异条带）。②转染哺乳动物细胞COS-7用夹心酶联免疫吸附法检测到了成纤维细胞生长因子18蛋白质的稳定表达（夹心酶联免疫吸附方法检测450nm处吸光度值，原液及1：10稀释度细胞上清大于阴性对照组2．1倍以上）。③用细胞上清刺激骨髓綦质干细胞后，实验组骨髓基质干细胞较对照组吸光度值明显增高，在24-72h其增高具有明屁的统计学差异（P〈0．01）。 结论：成纤维细胞生长因子18显著促进兔骨髓基质于细胞体外增殖，提示成纤维细胞生长因子18对于骨髓基质干细胞增殖及分化有调节作用。

英文摘要：

AIM： To construct stable eukaryotic expression plasmids of fibroblast growth factor-18 protein, and observe the in vitro effects of fibroblast growth factor-18 on the proliferation of rabbit bone marrow stem cells in vitro. METHODS： The experiment was conducted in the Military Institute of Bone Tumor and Stomatolotical Medical College of the Fourth Military University of Chinese PLA from October 2002 to March 2004. （1）The pGEM-T-fibroblast growth faetor-18-3 and pSecTag 2B expression vector were treated by enzyme restriction, and the pSecTag2B-fibroblast growth factor-18 plasmids were reconstructed. （2）The mammalian cell COS-7 was transfected under the mediation of bangosome, so as to stinmlate the bone marrow stem cells with cellular supcrrtatant following stable expression of COS-7. （3） MTT method was used to detect the proliferation of cells. RESULTS： （1）The eukaryotic expression plasmids of pSecTag2B-fibroblast growth factor-18 was constructed successfully （540bp special strip could be discerned in electrophoresis picture）. （2）The COS-7 was detected by ELISA method and stable expression of fibroblast growth factor-18 was detected （The absorbance value at 450 nm was detected by ELISA, and the it was 2.1 times of the original fluid and 1：10 dilution fluid as that in the negative control group）. （3） After the bone marrow stem cells were stimulated by cellular supernatant, the absorbance of bone marrow stem cells in the experiment group was significantly increased than that in the control group. There were statistical differences in the increased between the 24^th hour and the 72^nd hour, which were significant. CONCLUSION： Fibroblast growth factor-18 can significantly promote the proliferation of rabbit bone marrow stem cells in vitro, which suggests that the fibroblast growth factor-18 has regulative effect on the proliferation and differentiation of bone marrow stem cells.