华西口腔医学杂志

• 基础研究 • 上一篇    下一篇

大鼠牙囊细胞体外培养技术的建立及其鉴定

凌均棨,谷海晶,高燕,王阿丹   

  1. 中山大学光华口腔医院 牙体牙髓科,广东 广州510055
  • 收稿日期:2004-02-25 修回日期:2004-02-25 出版日期:2004-02-20 发布日期:2004-02-20
  • 通讯作者: 谷海晶,Tel:020-83822804
  • 作者简介:凌均棨(1953-),女,四川人,教授,博士

Establishment of Cultivating Technology of Dental Follicle Cells of Rat and Their Identifying

LINGJun-qi,GUHai-jing, GAO Yan,WANG A-dan   

  1. Dept.of Operative and Endodontics,Guanghua College of Stomatology,Sun Yat-Sen University, Guangzhou510055,China
  • Received:2004-02-25 Revised:2004-02-25 Online:2004-02-20 Published:2004-02-20

摘要:

目的 建立体外培养大鼠牙囊细胞的方法,观察牙囊细胞体外生长的生物学特性。方法 分离乳鼠下颌第一、第二磨牙牙胚的牙囊组织,应用组织培养的方法进行牙囊细胞原代和传代培养。显微镜观察培养细胞的形态,电镜观察体内牙囊组织和体外培养的牙囊细胞的超微结构。免疫组化法检测细胞波形蛋白、Ⅰ型胶原、纤维连接蛋白的表达。结果 分离组织培养24 h后,梭形细胞和多角形细胞从组织块中游出,经消化排除法获得纯化的牙囊细胞。显微镜下牙囊细胞为成纤维细胞样,电镜下牙囊细胞无桥粒,胞浆中含有高密度电子颗粒和大量粗面内质网(RER)。免疫组化染色显示牙囊细胞表达波形蛋白、Ⅰ型胶原和纤维连接蛋白。结论 运用组织培养和细胞生物学技术可以成功获得原代和传代培养的大鼠牙囊细胞,体外培养的细胞具有与正常大鼠牙囊细胞相一致的生物学特性。

关键词: 牙囊, 细胞培养, 成纤维细胞, 牙齿萌出

Abstract:

Objective To establish a method for culturing dental follicle cells of rat and observe the growth characteristics of the cultured cellsin vitro.Methods Dental follicle and associated enamel organs were dissected from the first and second man- dibular molars of 6-7-day-old rats, and then culturedin vitro. Purified dental follicle cells derived fromthe third orthe fourth pas- sage cells were utilized in the following experiments. The shape and ultrastructure of dental follicle cellswere observed by light-mi- croscopy and transmission electron microscopy. Immunocytochemistrywas used to detect the expression of vimentin, typeⅠcolla- gen and fibronectin.Results The cultured cells were fibroblastic in shape, elongate with processes, and transmission electron microscopy revealed that they contained electron-dense granules, an abundant rough endoplasmic reticulum, but did not form desmosomes. Vimentin, typeⅠcollagen and fibronectinwere present in dental follicle cell.Conclusion The dental follicle cells of rat could be successfully culturedin vitroand the cultured cells had the same characteristics of dental follicle cells of normal rat.

Key words: dental follicle, cell culture, fibroblasts, tooth eruption