MC3T3-E1前成骨细胞培养时间对矿化结节表达的影响

目的:观察MC3T3-E1前成骨细胞不同培养时间点矿化结节的形态,探讨一个既节省实验时间与经费,又便于观察矿化结节形态差异的实验方法。方法:将MC3T3-E1前成骨细胞按培养时间分为四组(14、21、28、35天组),各组实验结束时行茜素红染色,光学显微镜下观察矿化结节的形态变化。结果:各组均见红色的矿化结节形成,随培养时间延长,染色面积增大,密度增高,14天时结节轮廓清晰,结节间距较大,21天时结节面积增大,28天时结节边界超出视野,35天时视野内大片深染,结节轮廓不清。结论:在本实验周期内,MC3T3-E1前成骨细胞培养14至21天通过茜素红染色可以较清晰地观察矿化结节,其中培养14天时即可观察到结节大小、数量及形态,考虑到实验时间及经费的因素,我们认为MC3T3-E1前成骨细胞培养14天后行茜素红染色是观察不同因素对其矿化产生影响的适宜时间点。

Effects of Culture Time of MC3T3-E1 Pre-osteoblasts on the Expression of Mineralized Nodules

Objective:To observe the effect of MC3T3-E1 pre-osteoblasts at different culture time on mineralized nodules morphology, and explore an experimental method which not only can save experiment time and funds, but also is convenient to observe the morphology differences of mineralized nodules.Methods:The MC3T3-E1 pre-osteoblasts were divided into four groups according to the cultured time (14, 21, 28, 35 days group), then they were stained by alizarin red staining at the end of the experiment, to observe the morphological changes of the mineralized nodule under optical microscope.Results:The formation of mineralized nodule was detected in all the four groups. With the extension of the cultured time, the staining area increased, and the density also increased. On the 14th day, the nodules were clear, and the distance between nodules was large, and on the 21st day, the area of nodules increased, and on the 28th day, the nodule boundary were beyond vision, while on the 35th day, the field was largely hyperchromatic, and the outline of nodule was unclear.Conclusion:In the period of this experiment, it was suitable for MC3T3-E1 pre-osteoblasts to culture for 14 to 21 days by alizarin red staining. Cultured for 14 days, the cell number, size and morphology can be clearly observed when it is the best time to study the effects of different factors on the MC3T3-E1 pre-osteoblast mineralization.