模拟失重骨细胞条件培养基对成骨细胞活性的影响

发布时间：2019-02-27 20:29

【摘要】：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(P0.001);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。
[Abstract]:Objective to investigate the regulation of osteoblast activity by (RCM), a three-dimensional random rotating simulated weightlessness culture medium. Methods three-dimensional random rotation simulated weightlessness was used to culture mouse bone cell line MLO-Y448h. The conditioned medium (RCM) was collected and the conditioned medium (CCM), of the control group was collected by (MTT) method with tetramethylazo salt colorimetric method. The effects of RCM on the proliferation, cell cycle and alkaline phosphatase (ALP) activity of mouse osteoblast-like cell line MC3T3-E1 were detected by (pNPP) assay and flow cytometry (FCM). Results the proliferation of MC3T3-E1 was promoted by MLO-Y4RCM after 48 hours of three-dimensional random rotation. After MC3T3-E148h and 72 h culture with 100% MLO-Y4RCM and CCM, RCM group increased 1.21 and 1.27 times than CCM group, respectively, with significant difference (P0.001). The results of periodic detection showed that RCM could cause MC3T3-E1 to cross the periodic block point (S phase), and the ALP activity of MC3T3-E1 was not different between RCM group and CCM group. Conclusion the conditioned medium of three-dimensional random rotation simulated weightlessness culture for 48 hours can promote the proliferation of osteoblasts and make the osteoblasts cycle beyond the blocking point and have no effect on the activity of ALP of osteoblasts.
【作者单位】：西北工业大学生命科学院空间生物实验模拟技术国防重点学科实验室西北工业大学特殊环境生物物理学研究所;
【基金】：国家自然科学基金(30700152)资助
【分类号】：R851.22

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