小鼠单核细胞RAW264.7的细胞生物学特征

Biological Characteristics of Mice Monocyte Cell RAW264.7

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.