解离的小鼠牙胚上皮细胞在重聚发育过程仍维持牙齿发育基因的表达.PDFVIP

生 物 工 程 学 报 Chin J Biotech 2010, December 25; 26(12): 1690−1695 journals.im.ac.cn Chinese Journal of Biotechnology ISSN 1000-3061 cjb@im.ac.cn ©2010 CJB, All rights reserved. 组织工程与细胞培养 解离的小鼠牙胚上皮细胞在重聚发育过程仍维持牙齿发 育基因的表达 胡雪峰，林陈胜，王冰梅，韩萍萍，张彦定 福建师范大学生命科学学院 福建省发育与神经生物学重点实验室，福州 350108 摘 要: 为了解牙胚细胞解离重聚过程的细胞形态和分子机制，将小鼠帽状期牙胚解离细胞重聚，移植到小鼠肾囊膜 下培养，组织切片，HE 染色，观察再生牙齿的形态发生过程，并用原位杂交的方法进一步检测了与牙上皮发育密切相 关的基因在再生牙上皮中的表达情况。结果发现，解离重聚的牙胚细胞在牙齿器官的再生过程重现了正常牙齿的形态 发生过程；解离的牙上皮细胞在重聚和再生过程中保持Fgf8 、Noggin 和Shh 等牙上皮发育基因表达。以上结果表明， 即便是被解离形成分散状态的牙上皮细胞，在与牙胚间充质细胞重新聚合后，仍保持牙向分化的潜能。该结果为理解 牙齿再生的机理提供新的实验数据，对利用干细胞进行牙齿再生的研究有重要的提示意义。 关键词: 牙齿再生，细胞重聚，牙上皮，基因表达 Dissociated mouse tooth germ epithelial cells retain the expression of tooth developmental genes during reaggregation process Xuefeng Hu, Chensheng Lin, Bingmei Wang, Pingping Han, and Yanding Zhang Key Laboratory of Developmental and Neuro Biology of Fujian Province, College of Life Sciences, Fujian Normal University, Fuzhou 350108, China Abstract: Generation of bio-engineered teeth by using stem cells will be a major approach for bioengineered implantation. Previous studies have demonstrated that dissociated tooth germ cells are capable of generating a tooth after reaggregation in vitro. However, the cellular and molecular mechanisms underlying this tooth regeneration are not clear. In this study, we dispersed E13.5 molar germ into single cells, immediately reaggregated them into cell pellet, then grafted the reaggregates under mouse kidney capsule for various times of culture. We investigated the morphogenesis and the expression of several developmental genes in dental epithelial cells in reaggregates of tooth germ cells. We found that dissociated tooth germ c