建立小鼠成骨与破骨细胞相互作用的共育新体系

背景：众所周知，骨重建是骨组织中重要的生物学反应过程，其中成骨细胞与破骨细胞发挥了关键作用。但目前，关于骨重建中成骨与破骨细胞间信号传递的深层机制还不清楚。 目的：利用transwell技术，在体外建立一种成骨与破骨细胞的新型共育体系，为深入研究骨重建中成骨与破骨细胞的相互作用提供成熟的实验模型。 方法：采用MC3T3-E1成骨样细胞株与RAW264.7破骨前体细胞株，进行体外成骨与破骨细胞的诱导分化，并利用Transwell共培养板(0.4 µm聚酯膜)建立成骨与破骨细胞的共育体系。共培养6 d后，通过测定细胞活性和碱性磷酸酶(ALP)活力分析成骨细胞的增殖和分化活性，利用抗酒石酸酸性磷酸酶(TRAP)染色、甲苯胺蓝(TB)染色、TRAP活性测定及扫描电镜技术观察破骨细胞的分化及骨吸收功能。 结果与结论：共培养体系中成骨样细胞的无限增殖能力减弱，而分化活性明显增强，同时破骨前体细胞被诱导分化为成熟的破骨细胞，并具有一定的骨吸收功能。因此，该共培养体系可用于骨重建中成骨与破骨细胞间信号通路的深层研究。

A novel co-culture system for interaction of mouse osteoblasts and osteoclasts

BACKGROUND:It is well known that bone remodeling is an important biological process in which osteoblasts and osteoclasts play the critical role. However, the detailed mechanisms of osteoblast-osteoclast communication in bone remodeling remain unclear. OBJECTIVE:To in vitro establish a novel co-culture system for interaction of mouse osteoblasts and osteoclasts using the Transwell technology. METHODS:Osteoblastic cell line MC3T3-E1 and preosteoclasts RAW264.7 were selected, and then the cells were induced to differentiate into osteoblasts and osteoclasts, respectively. The co-culture system for osteoblasts and osteoclasts interaction was established with Transwell co-culture plate (0.4 µm polyester film), and the osteoblasts and osteoclasts were co-cultured for 6 days. After that, the proliferation and differentiation activities of osteoblasts were analyzed through testing the cell activity and alkaline phosphatase activity, and the differentiation and bone resorption of osteoclasts were observed using tartrate-resistant acid phosphatase staining, toluidine blue staining, tartrate-resistant acid phosphatase activity detecting and scanning microscope technology.   RESULTS AND CONCLUSION:In the co-culture system, the unlimited proliferation ability of osteoblasts was decreased, while the differentiation activity was increased. Meanwhile, the preosteoclasts could differentiate into mature osteoclasts with bone resorption function. Thus, this co-culture system can be applied in the further study of osteoblast-osteoclast communication in bone remodeling.