大鼠牙囊细胞体外培养与表型鉴定  

The culture and phenotype identification of rat dental follicle cells in vitro

作　　者：徐璐璐[1] 刘洪臣[2] 王东胜[2] 鄂玲玲[2] 

机构地区：[1]解放军总医院口腔正畸科,主治医师北京100853 [2]解放军总医院口腔医学中心,主任主任医师教授北京100853

出　　处：《中华老年口腔医学杂志》2010年第2期98-101,共4页Chinese Journal of Geriatric Dentistry

基　　金：国家自然科学基金资助项目(30400510)

摘　　要：目的:体外培养大鼠牙囊细胞(RDFCs),鉴定其细胞来源及表型,为牙周组织再生的研究提供可靠的种子细胞来源。方法:选择出生后6-7d SD仔鼠,分离上、下颌第一、第二磨牙牙胚,取牙囊组织进行原代和传代培养。通过倒置显微镜观察细胞形态及生长情况;波形丝蛋白(Vimentin,VIM)和角蛋白(Cytokeratin,CK)鉴定其细胞来源;免疫组化染色技术检测细胞骨桥蛋白(osteopontin,OPN)、骨涎蛋白(bone sialoprotein,BSP)、Ⅰ型胶原(type Ⅰ collagen,CoL-Ⅰ)、Ⅲ型胶原(typeⅢ collagen,CoL-Ⅲ)及纤维连接蛋白(fibronectin,FN)的表达。结果:细胞形态呈多形性,有长梭形,纺锤形、不规则三角形等,波形丝蛋白染色阳性,角蛋白染色阴性。免疫组化染色显示OPN、BSP、CoL-Ⅰ、CoL-Ⅲ、FN在胞浆中呈不同程度的阳性表达。结论:大鼠牙囊细胞来源于外胚间充质,具有成纤维细胞的形态特征及成骨/成牙骨质细胞表型特征,可将牙囊细胞作为种子细胞用于牙周组织工程的再生研究。Objective：To culture the rat dental follicle cells in vitro and to identify its origin. Meanwhile,to investigate them as seeded cell for periodontal tissue engineering. Methods：Dental follicle and associated enamel organs were dissected from the first and second molars of 6 or 7-day-old rats,and then cultured in vitro. Vimentin and cytokeratin were examined to identify its origin. Immunocytochemistry was used to detect the expression of osteopontin,bone sialoprotein,collagen type I,collagen type Ⅰ and fibronectin. Results：The cultured RDFCs showed multiformity,elongate shape,spindle shape,and irregular triangle shape,etc. The expression of vimentin was positive while that of cytokeratin was negative. Osteopontin,bone sialoprotein,collagen type I,collagen type Ⅰ and fibronectin were expressed in the RDFCs. Conclusions：The results indicated that the RDFCs could be cultured in vitro and these cells showing multiformity originate from ectomesenchyme. RDFCs could be used as the seeded cells for periodontal tissue engineering.

关 键 词：牙囊细胞 细胞培养 鉴定 

分 类 号：R783[医药卫生—口腔医学]