Han Xue, Yoshizaki Keigo, Tian Tian, Miyazaki Kanako, Takahashi Ichiro, Fukumoto Satoshi
Section of Orthodontics and Dentofacial Orthopedics, Division of Oral Health, Growth and Development, Kyushu University Faculty of Dental Science, Fukuoka, Japan.
Division of Pediatric Dentistry, Department of Oral Health and Development Sciences, Tohoku University Graduate School of Dentistry, Sendai, Japan.
Bio Protoc. 2020 Feb 5;10(3):e3515. doi: 10.21769/BioProtoc.3515.
A tooth germ organ culture allows visualization of its development in different stages, thus enabling investigation of the molecular mechanisms of regulatory factors. Tooth germs can be rapidly dissected from E13 mouse embryos and placed on cell culture inserts for observation of subsequent tooth germ development in a three-dimensional situation in real time. This method is also suitable for other organs that develop by epithelial-mesenchymal interactions, including salivary gland, hair, lung, and kidney. In addition, siRNAs or growth factors can be easily added to tooth germ cultures to investigate the detailed molecular function of specific genes. The present protocol provides an efficient and practical method for isolation and culture of embryonic tooth germs.
牙胚器官培养能够观察其不同阶段的发育过程，从而有助于研究调控因子的分子机制。牙胚可从E13小鼠胚胎中快速分离出来，并置于细胞培养插入物上，以便实时观察三维环境中牙胚的后续发育情况。该方法也适用于其他通过上皮-间充质相互作用发育的器官，包括唾液腺、毛发、肺和肾脏。此外，可轻松地将小干扰RNA（siRNAs）或生长因子添加到牙胚培养物中，以研究特定基因的详细分子功能。本实验方案提供了一种高效实用的胚胎牙胚分离和培养方法。
