摘要
目的利用双荧光标记小鼠模型观察破骨细胞的融合过程并探讨不同浓度的细胞核因子κB受体活化因子配体(RANKL)和甲状旁腺激素(PTH)对破骨细胞融合效率的影响。方法使用5周龄CTSK-Cre和的ROSAmt/mg两种转基因小鼠,前者无荧光表达而在其破骨细胞中表达CRE重组酶,后者表达红色荧光(tomato)。处死两种小鼠并冲出骨髓间充质干细胞(BMSCs)且以1∶1的比例混合培养。将不同浓度的RANKL和PTH加入共培养体系,计算绿色荧光的量。结果 CTSKCre和ROSAmt/mg2种转基因小鼠BMSCs共培养第5天观察到绿色荧光阳性的细胞出现;RANKL组:随着RANKL浓度的升高,共培养体系内绿色荧光阳性的细胞数不断增加;PTH组:共培养体系内绿色荧光阳性的细胞数与PTH浓度之间未发现明显的相关性。结论基于cre-loxp系统建立的cre重组酶诱导的双荧光标记小鼠模型可有效用于破骨细胞融合过程的研究;RANKL的浓度越高其诱导破骨细胞融合的效率越高;PTH与破骨细胞的融合效率之间无明显的剂量相关规律。
Objective To investigate the process of osteoclasts fusion by using a double fluorescent labeled mice mode and the effect of different concentrations of receptor activator of NF-kB ligand （RANKL） and parathyroid hormone （PTH） on the fusion rate. Methods We adopted two kinds of transgenic mouse, one was CTSK-Cre with non-fluorescence, and another was ROSAmt/mg, which can express tomato. The two types of mice were sacrificed and bone mesenchymal stem cells （BMSCs） were obtained. The two types of BMSCs were co-cultured at a ratio of 1 ： 1. In order to investigate the effect of different factors on osteoclast fusion, we added different concentrations of RANKL or PTH to the co-cuhure system, and then calculated the amount of green fluorescence （GFP）. Results Af- ter 5 days of the co-culture, there were GFP positive cells. In the RANKL group, the GFP positive cells were gradually increased with in- crease of RANKL concentrations. In the PTH group, there was no significant relationship between the concentrations of PTH and the numbers of GFP positive cells. Conclusion The high concentration of RANKL can induce a high efficiency of osteoclasts fusion, but not PTH.
出处
《实用医院临床杂志》
2017年第5期20-22,共3页
Practical Journal of Clinical Medicine
