牙源性角化囊肿上皮细胞体外培养及其生物学特性  

IN VITRO CULTURE AND BIOLOGICAL CHARACTERISTICS OF EPITHELIAL CELLS IN ODONTOGENIC KERATOCYST

作　　者：杨绍滨 张静[2] 薛娇 庞宝兴[4] 袁荣涛[1,3] YANG Shaobin;ZHANG Jing;XUE Jiao;PANG Baoxing;YUAN Rongtao(College of Stomatology,Qingdao University,Qingdao 266003,China)

机构地区：[1]青岛大学口腔医学院,山东青岛266003 [2]青岛市市立医院手术室 [3]青岛市市立医院口腔医学中心 [4]青岛大学附属医院口腔颌面外科

出　　处：《精准医学杂志》2019年第4期345-349,共5页Journal of Precision Medicine

基　　金：山东省自然科学基金面上项目(ZR2016HM34)

摘　　要：目的探讨体外培养牙源性角化囊肿上皮细胞的方法,并且比较牙源性角化囊肿和正常口腔黏膜(NOM)上皮细胞增殖能力的差异,为牙源性角化囊肿的体外实验提供细胞模型。方法采用酶消化法,在角化细胞无血清培养基上原代培养牙源性角化囊肿和NOM上皮细胞。倒置显微镜下观察细胞形态学特征,免疫荧光染色鉴定细胞表面标记物,并通过生长曲线、克隆形成率和细胞周期分析比较两种细胞的增殖能力差异。结果体外培养的牙源性角化囊肿上皮细胞表现为典型的上皮细胞铺路石样外观,体外培养可传至3~4代。经过CK、CK10、CK14、Vimentin免疫荧光染色鉴定为角化上皮细胞。牙源性角化囊肿上皮细胞在生长速度、克隆形成率和增殖指数等方面均高于NOM上皮细胞。结论采用酶消化法在角化细胞无血清培养基上可实现牙源性角化囊肿上皮细胞体外连续培养,牙源性角化囊肿上皮细胞较NOM上皮细胞具有更强的增殖活力。[Objective]To investigate the method for in vitro culture of odontogenic keratocyst(OKC)epithelial cells and the difference in proliferative capacity between OKC and normal oral mucosal(NOM)epithelial cells,and to provide a cell model for in vitro experiment of OKC.[Methods]Enzyme digestion was used for the primary culture of OKC and NOM epithelial cells in serum-free medium for keratinocytes.An inverted microscope was used to observe cell morphology;immunofluorescent staining was used to identify cell surface markers;growth curve,cloning efficiency,and cell cycle analysis were used to compared the diffe-rence in proliferative capacity between the two types of epithelial cells.[Results]OKC epithelial cells cultured in vitro had the typical cobblestone appearance of epithelial cells and were passaged to the third or fourth generation in vitro.They were identified as keratinized epithelial cells by immunofluorescence staining of CK,CK10,CK14,and vimentin.OKC epithelial cells had higher growth rate,cloning efficiency,and proliferation index than NOM epithelial cells.[Conclusion]Enzyme digestion can achieve continuous in vitro culture of OKC epithelial cells in serum-free medium for keratinocytes,and OKC epithelial cells have higher proliferative ability than NOM epithelial cells.

关 键 词：牙源性囊肿 上皮细胞 原代细胞培养 细胞增殖 

分 类 号：R739.82[医药卫生—肿瘤]