骨样细胞MLO-Y4与成骨样细胞MC3T3-E1生物学特性的比较

The Comparison of Biological Characteristics between Osteocyte-like Cell MLO-Y4 and Osteoblast-like Cell MC3T3-E1

分别采用倒置显微镜观察法、细胞计数法、RT-PCR法、磷酸对硝基苯酚法(PNPP法)和ELISA法来比较小鼠骨样细胞MLO-Y4与小鼠成骨样细胞MC3T3-E1的细胞形态、增殖、相关基因的表达和分泌功能的差异。结果显示MC3T3-E1细胞呈长梭形,具有少量短的突触;而MLO-Y4细胞呈星状或树枝状且具有很多长的突触。MC3T3-E1细胞的增殖能力强于MLO-Y4细胞,两者的倍增时间分别是18 h和20 h。MC3T3-E1细胞中原癌基因c-fos和骨桥蛋白基因OPNmRNA的表达明显高于MLO-Y4细胞,而骨钙素基因OCmRNA的表达则是MC3T3-E1细胞远低于MLO-Y4细胞,白细胞分化抗原44基因CD44 mRNA在两种细胞中的表达差异不明显。ALP的分泌在MC3T3-E1细胞中高于MLO-Y4细胞,NO的分泌在两种细胞中没有显著性差异,M-CSF在MLO-Y4细胞中的分泌较高。由此可见骨样细胞MLO-Y4与成骨样细胞MC3T3-E1在形态、ALP和MCSF分泌及c-fos、OPN和OCmRNA表达方面差异明显。

To compare the biological characteristics of osteocyte-like cell MLO-Y4 and osteoblast-like cell MC3T3-E1 from cell morphology, cell proliferation, cell secretion and gene expression with different methods： Cell morphology was observed by inverted microscope. Cell counting assay was used to detect cell proliferation. RT- PCR was used to detect mRNA expression of OC, CD44, OPN and c-fos. Secretion of ALP was measured by PNPP. Secretion of M-CSF and NO were measured by ELISA. The results tell us： osteocyte-like cell MLO-Y4 and osteoblast-like cell MC3T3-EI showed obvious difference in cell morphology, MC3T3-EI cells were spindle-shaped with few short dendrites, however, MLO-Y4 cells were star-shaped or dendrite-shaped with more dendrites. The doubling time of MC3T3-E1 cells was 18 h, while that of MLO-Y4 cells was 20 h. RT-PCR showed the expression of c-fos and OPN were higher and the expression of OC was lower in MC3T3-E1 cells than in MLO-Y4 cells, and there was not significant difference of CD44 expression between the two kinds of cells. ALP secretion was higher and M- CSF secretion was lower in MC3T3-E1 cells than that in MLO-Y4 ceils. There is not significant difference of NO secretion between the two kinds of cells. It can be seen that there were significant differences in cell morphology, ALP and M-CSF secretion, expression of c-fos, OPN and OC between MLO-Y4 cells and MC3T3-Elcells. In bone research MLO-Y4 cells and MC3T3-E1 cells are often used. According to their different biological characteristics, researchers could choose appropriate cell types to in their study.