人牙囊细胞的分离培养和生物学特性

Cultivating human dental follicle cells in vitro and their characteristics

目的 :应用酶消化法体外培养人牙囊细胞并研究其生物学特性。方法 :分离合法引产人胚胎乳牙胚的牙囊组织 ,消化法获得人牙囊细胞 ,HE染色观察细胞形态、免疫组织化学染色技术检测波形丝蛋白、Ⅰ型胶原表达 ,RT -PCR检测细胞的骨钙素、骨涎蛋白、碱性磷酸酶的表达。结果 :原代培养的人牙囊细胞呈成纤维细胞形态 ,有部分上皮成分混杂 ,经纯化后可排除 ;在牙囊细胞中Ⅰ型胶原和波形丝蛋白的表达呈阳性 ,定位于细胞的胞质中 ;RT -PCR结果显示骨涎蛋白、骨钙素、碱性磷酸酶表达。结论 :体外培养的人牙囊细胞具有成纤维细胞的特性 ,具有合成Ⅰ型胶原的能力 ;不同程度表达Ⅰ型胶原、骨钙素、骨涎蛋白、碱性磷酸酶等矿化相关蛋白。

AIM:To culture human dental follicle cells (HDFC) in vitro and study their characterization.METHODS:Dental follicle tissue was dissected from the root surface of human embryo tooth germs and cells were released by digestion with bacterial collagenase.Observing the morphology by microscope and HE staining.Identifying the cells′ characterization by immunohistochemistry and RT-PCR method.RESULTS:The cultured human dental follicle cells in vitro showed fibroblasts morphology.The immunohistochemistry staining showed that the expression of type Ⅰcollagen and vimentin was positive and localized in the cytoplasma.CONCLUSION: The mRNA of OCN,BSP and ALP associated with mineralization was also detected by RT-PCR technique.