In vitrothree-dimensional development of mouse molar tooth germs in a rotary cell culture system


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Abstract

Objective.In vitro tooth germ cultivation is an effective method to explore the mechanism of odontogenesis. The three-dimensional rotary cell culture system (RCCS) is typically used to culture simulated organs such as cartilage, skin, and bone. In this study, we established an in vitro tooth germ culture model using RCCS to investigate whether RCCS could provide an appropriate environment for tooth germ development in vitro.Methods.Mandibular first molar tooth germs from 1-day post-natal mice were cultured in RCCS for 3, 6, and 9 days. Tooth germ development was monitored via histology (hematoxylin & eosin staining), stereoscopic microscopy, and quantitative real-time PCR (RT-PCR).Results.Tooth germs cultured in RCCS maintained their typical spatial shape. Blood vessels were maintained on the dental follicle surface surrounding the crown. After cultivation, thick layers of dentin and enamel were secreted. Compared with tooth germs grown in jaw, the tooth germs grown in RCCS exhibited no significant difference in DMP1 or FGF10 expression at all time points.Conclusions.Use of RCCS enhanced the development of tooth germs and allowed the tooth germs to maintain their spatial morphology. These results indicate that RCCS may be an effective culture system to investigate the mechanism of tooth development.International Journal of Paediatric Dentistry 2014; 24: 175–183

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