犬根尖牙乳头干细胞的分离培养和生物学特性  

Isolation,culture and biological characterization of Beagle stem cells from apical papilla

作　　者：杨杰[1] 赵玉鸣[1] 王文君[1] 贾维茜[1] 葛立宏[1] 

机构地区：[1]北京大学口腔医学院.口腔医院儿童口腔科,北京100081

出　　处：《北京大学学报（医学版）》2012年第6期921-926,共6页Journal of Peking University:Health Sciences

基　　金：国家自然科学基金(81170928)资助~~

摘　　要：目的:分离培养比格犬(Beagle犬)根尖牙乳头干细胞并研究其生物学特性。方法:拔除Beagle犬上颌年轻恒前牙,切取根尖牙乳头,采用酶消化法分离培养Beagle犬根尖牙乳头细胞,进行成纤维细胞克隆形成实验、生长曲线测定、多向分化能力研究及间充质干细胞相关表面标记物STRO-1、CD146和CK的表达分析,并将其移植至免疫缺陷鼠皮下观察矿化组织的形成。结果:Beagle犬根尖牙乳头组织中存在一群具有高度增殖能力的细胞,在相同培养条件下,其克隆形成率明显高于人根尖牙乳头干细胞,差异具有统计学意义(P<0.001);所获得Beagle犬根尖牙乳头干细胞具有成骨、成脂和成软骨等多向分化能力;在体外经成骨诱导后可形成矿化结节,成脂诱导后可形成脂滴,成软骨诱导后免疫组织化学染色Ⅱ型胶原阳性表达;同时该群细胞STRO-1和CD146阳性表达,而CK阴性表达;与羟基磷灰石混合移植于免疫缺陷鼠皮下可形成矿化组织和牙髓-牙本质复合体样结构。结论:Beagle犬根尖牙乳头中存在具有高增殖能力和多向分化能力的根尖牙乳头干细胞。Objective：To isolate and characterize the Beagle stem cells from apical papilla.Methods：Apical papilla was severed from the end of freshly extracted Beagle＇s young permanent upper anterior teeth,and digested by collagenase type Ⅰ and dispase for cell culture.The isolated cells were investigated for stem cell properties by analyzing their colony-forming efficiency,growth characteristics and the expression of mesenchymal stem cell markers;and evaluating their multidifferentiation potentials including osteogenic,adipogenic,and chondrogenic potentials.Additionally,the cells were transplanted subcutaneously into immunocompromised mice to observe the mineral tissue formation.Results：Our study showed that a clonogenic,rapidly proliferative population of cells existed in Beagle＇s apical papilla,and these cells had a significantly higher colony-forming rate than the stem cells from apical papilla derived from humans（P0.001）.These cells had multilineage differentiation ability including osteogenic,adipogenic and chondrogenic potentials.Mineralized nodules were formed after osteogenic induction,lipid droplets were found after adipogenic induction,and the pellets showed positive immunohistochemical staining for collagenⅡafter chondrogenic induction.These cells also expressed the mesenchymal stem cell markers including STRO-1 and CD146,while negative for CK.Moreover,these cells transplanted with hydroxyapatite in immunocompromised mice could form mineral tissue and pulp-dentin complex-like tissue.Conclusion：There are stem cells from the apical papilla which have high proliferation ability and multilineage differentiation potential existing in the Beagle＇s apical papilla.

关 键 词：根尖牙乳头干细胞 细胞增殖 多向分化 犬 

分 类 号：R322.4[医药卫生—人体解剖和组织胚胎学]