华西口腔医学杂志

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小鼠牙囊细胞的体外培养及表型特征

葛少华,杨丕山,赵 宁,李 纾   

  1. 山东大学口腔医院 牙周病科,山东 济南250012
  • 收稿日期:2005-10-25 修回日期:2005-10-25 出版日期:2005-10-20 发布日期:2005-10-20
  • 通讯作者: 杨丕山, Tel:0531-88382368
  • 作者简介:葛少华(1973-),女,山东人,讲师,硕士
  • 基金资助:

    家自然科学基金资助项目(30371537);山东省优秀中青年科学家奖励基金资助项目(02BS096)

Culture and Phenotype Characteristics of Mouse Dental Follicle Cells

GE Shao-hua,YANG Pi-shan,ZHAO Ning,LI Shu   

  1. Dept.ofPeriodontology,School ofStomatology,Shandong University,Jinan250012,China
  • Received:2005-10-25 Revised:2005-10-25 Online:2005-10-20 Published:2005-10-20

摘要:

目的 建立小鼠牙囊细胞(DFC)的体外分离培养方法,研究牙囊细胞的表型特征。方法 用1%的胰蛋白酶消化分离9 d龄Balb/c小鼠的下颌第一磨牙牙胚的牙囊组织进行体外培养,倒置显微镜下观察细胞的形态,扫描电镜观察细胞的表面形貌,SP免疫组化法检测细胞碱性磷酸酶(ALP)、骨涎蛋白(BSP)、骨桥蛋白(OPN)的表达。结果 分离培养的细胞有3种类型:立方形或多角形;长梭形;非常细长的细胞形状。扫描电镜下细胞具有多形性,有大量的线状胞浆突和微绒毛;根据细胞表面有无纤维细丝覆盖,可将细胞分为有纤维细丝、无纤维细丝两个亚群。免疫组化染色显示部分牙囊细胞的ALP、BSP、OPN的表达阳性。结论 牙囊细胞具有多种细胞的表型特征, 有分化为成牙骨质细胞、牙周膜细胞和成骨细胞的潜能。

关键词: 牙囊, 碱性磷酸酶, 骨桥蛋白, 骨涎蛋白

Abstract:

Objective To establish a method for isolating and culturing mouse dental follicle cells and to study the phenotype characteristics of dental follicle cells.Methods Mandibular first molars from 9 day old Balb/c mice were digested with 1% tryp- sin, subsequently, and the dental follicle was enucleated from the tooth germ and cultured. The shape and ultrastructural appear- ance of dental follicle cellswere observed under phase-contrastmicroscope and scanning electron microscope(SEM). Immunocyto- chemistrywas used to detect the expression of alkaline phosphatase (ALP), bone sialoprotein(BSP) and osteopontin(OPN).Re- sults Three types of cells were isolated: some were cuboidal/polygonal; some were elongated, spindle-shaped, fibroblast-like cells; and a minor, third cell type was very thin and elongated. The cytoplasm of the first two cell types was filled with abundant granules. The cells were pleomorphism under SEM and had many filipodia and microvilli. According to whether there were fila- ments overlying the surface, the cells could be divided into two subtypes. Some but not all follicle cells expressed ALP, BSP and OPN.Conclusion The cultured dental follicle cells consisted of several cell phenotypes and had the potential of differentiating in- to cementoblasts, periodontal ligament fibroblasts and osteoblasts.

Key words: dental follicle, alkaline phosphatase, bone sialoprotein, osteopontin