Sun Liang, Yang Chao, Ge Yaneng, Yu Mei, Chen Guoqing, Guo Weihua, Tian Weidong
Regenerative Medicine Research Center, West China Hospital, Sichuan University, Chengdu, Sichuan, China; National Engineering Laboratory for Oral Regenerative Medicine, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China; State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China.
Int J Paediatr Dent. 2014 May;24(3):175-83. doi: 10.1111/ipd.12057. Epub 2013 Jul 19.
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.
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).
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.
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.
体外牙胚培养是探索牙发生机制的有效方法。三维旋转细胞培养系统（RCCS）通常用于培养软骨、皮肤和骨骼等模拟器官。在本研究中，我们使用RCCS建立了体外牙胚培养模型，以研究RCCS是否能在体外为牙胚发育提供适宜的环境。
将出生后1天小鼠的下颌第一磨牙牙胚在RCCS中培养3、6和9天。通过组织学（苏木精和伊红染色）、立体显微镜和定量实时PCR（RT-PCR）监测牙胚发育。
在RCCS中培养的牙胚保持其典型的空间形状。牙冠周围的牙囊表面有血管分布。培养后，分泌出厚层的牙本质和釉质。与在颌骨中生长的牙胚相比，在RCCS中生长的牙胚在所有时间点的DMP1或FGF10表达均无显著差异。
使用RCCS可促进牙胚发育，并使牙胚保持其空间形态。这些结果表明，RCCS可能是研究牙齿发育机制的一种有效培养系统。
