摘要
目的观察小鼠单核/巨噬细胞RAW264.7的一般细胞生物学特征。方法用倒置相差显微镜观察细胞的形态特征,胎盘蓝染色细胞计数,绘制细胞生长曲线。半定量逆转录聚合酶链反应观察细胞3~2l天破骨细胞表型和功能基因表达谱变化。结果RAW264.7细胞形态以类圆形和不规则多边形为主,一般含l~2个核,有伪足,细胞胞体较小。细胞生长快,5~6天可汇合。RAW264.7细胞能表达成熟破骨细胞的表型标志基因如核因子KB受体活化子(RANK)、抗酒石酸酸性磷酸酶(TRAP)、基质金属蛋白酶9(MMP9)及骨吸收有关的功能基因如组织蛋白酶K(CaK)、整合素av(integrin av)等。RAW264。7细胞培养3~21天,上述基因的mRNA表达不随培养时间改变。结论RAW264.7具有破骨细胞特征性基因表达谱,是一种较好的破骨前体细胞模型。
Objective To observe the general biological characteristics of mice monocyte/macrophage cell RAW264.7. Methods The morphology of RAW264.7 cells was observed by inverted phase contrast microscope, cells were counted using placenta blue-stain to protract cell growth curve. Gene expression of RAW264.7 from the 3rd to the 21st day was observed by RT-PCR. Results Cells were characterized by adherent appearance, with round-shaped and irregular polygon with 1 - 2 nuclei, pseudopodium and small cell body. Cells reached confluence in 5 - 6 day. Most of the osteoelastic phenotypes or functioning genes as MMP9, TRAP and RANK could be detected in RAW264.7 cells, indicating that it belongs to a kind of osteoclast precursor cells. The gene expression of RAW264.7 cells were subjected to no detected changes in cultures from 3 to 21 days. Conclusion RAW264.7 presents specific phenotypes of osteoelast and the expression of functioning genes is constant, showing a typical moldel of the osteoclast precursor cells.
出处
《南华大学学报(医学版)》
2008年第3期282-285,共4页
Journal of Nanhua University(Medical Edition)
基金
湖南省自然科学基金(OTJJ6160)
湖南省卫生厅课题(2007117)