Abstract:

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.

Key words: tissue construction, bone tissue construction, co-culture, osteoblasts, osteoclasts, alkaline phosphatase, tartrate-resistant acid phosphatase, National Natural Science Foundation of China