论文部分内容阅读
目的探讨三维随机回转模拟失重培养后的骨细胞条件培养基(RCM)对成骨细胞活性的调节作用。方法采用三维随机回转模拟失重培养小鼠骨细胞系MLO-Y448h后,收集回转条件培养基(RCM)和未回转对照组的条件培养基(CCM),用四甲基偶氮唑盐比色(MTT)法、对硝基苯磷酸(pNPP)法和流式细胞术(FCM)检测RCM对小鼠成骨样细胞系MC3T3-E1增殖、周期及细胞分泌碱性磷酸酶(ALP)活性的影响。结果三维随机回转48h后的MLO-Y4RCM可促进MC3T3-E1增殖,用100%的MLO-Y4RCM和CCM培养MC3T3-E148h和72h后,RCM组比CCM组分别增加了1.21和1.27倍,差异性极显著(P<0.001);周期检测结果表明,RCM可以使MC3T3-E1越过周期阻滞点(S期);MC3T3-E1的ALP活性在RCM组和CCM组之间无差异。结论三维随机回转模拟失重培养骨细胞48h后的条件培养基促进了成骨细胞的增殖;并使成骨细胞的周期越过阻滞点,对成骨细胞的ALP活性无影响。
Objective To investigate the regulation of osteoblast activity by osteoblast conditioned medium (RCM) after three-dimensional random rotation simulated weightlessness culture. Methods MLO-Y448h mice were cultured with three-dimensional random rotation to simulate weightlessness. Conditioned medium (CCM) was collected from rotating conditioned medium (RCM) and non-rotating control group, MTT assay, pNPP assay and flow cytometry (FCM) were used to detect the effects of RCM on the proliferation, cell cycle, and alkaline phosphatase (ALP) activity of mouse osteoblast-like cell line MC3T3-E1 . Results MLO-Y4RCM increased the proliferation of MC3T3-E1 after three-dimensional random revolving 48h. After MC3T3-E148h and 72h cultured with 100% MLO-Y4RCM and CCM, the RCM group increased 1.21 and 1.27 times respectively compared with CCM group, (P <0.001). The result of cycle test showed that RCM could make MC3T3-E1 cross the cycle arrest point (S phase). The ALP activity of MC3T3-E1 had no difference between RCM group and CCM group. CONCLUSIONS: Conditioned medium supplemented with three-dimensional random swift simulated weightlessness cultured osteoblasts stimulated the proliferation of osteoblasts, and the period of osteoblasts crossed the arresting point, which had no effect on the ALP activity of osteoblasts.