小鼠牙囊细胞体外培养方法与形态特征探讨  

Exploration of culture method and morphological characteristics of mouse dental follicle cells in vitro

作　　者：王贺[1,2,3] 吴补领 段小红[3] 

机构地区：[1]南方医科大学南方医院 [2]南方医科大学口腔医学院,广东广州510515 [3]军事口腔医学国家重点实验室陕西省口腔医学重点实验室第四军医大学口腔医院口腔生物学教研室,陕西西安710032

出　　处：《牙体牙髓牙周病学杂志》2015年第6期370-374,共5页Chinese Journal of Conservative Dentistry

基　　金：国家自然科学基金(81271116)

摘　　要：目的：：探讨小鼠牙囊细胞的体外培养方法并观察其形态特征。方法：体视显微镜下分离出生3~5d仔鼠牙胚,用I型胶原酶消化后分离牙囊组织,采用酶消化-组织块法和二次酶消化法进行牙囊细胞的原代及传代培养,倒置显微镜下进行细胞形态学观察。结果：酶消化-组织块法经4~5 d,部分组织块周围见细胞呈放射状爬出,2周后细胞集落仍难以汇合,无法传代。二次酶消化法第2天可见散在细胞集落形成,4~5d细胞接触汇合。传代发现,多角形细胞消失,第2代后梭形细胞占大多数,第3代细胞生长速度最快,第4代细胞增殖速度减慢,第5代后细胞趋于静止；随着传代次数的增加胞体逐渐增大,可出现一些树突形细胞。结论：采用二次酶消化法进行小鼠牙囊细胞原代培养可在短时间内获得较多的细胞,操作简便且成功率高,细胞形态随培养时间而变化多样。AIM：To explore the culture method of mouse dental follicle cells （ DFCs） in vitro and to observe their morphological characteristics. METHODS：Dental germs from 3-5 days postnatal mice were digested with type I collagenase and the dental follicle tissues were dissected by enzyme isolation-tissue explant and twice-enzyme isola-tion. The morphology of the cultured DFCs was observed under inverted microscope. RESULTS：At the 4 th-5 th day in vitro, several cells were extended from part of the tissue explants and were hardly to fuse even after 2 weeks′culture. The method of enzyme isolation-tissue explant was failed because of unable to passage and amplify the cells while the method of twice-enzyme isolation got a success rate of about 85%. Scattered groups of cells were seen at the second day and they fused at the 4 th-5 th day in vitro by way of twice-enzyme isolation. Polygonal and spindle cell populations co-existed in primary culture. Almost all polygonal cells disappeared in subculture, and spindle cells became the majority of cell population. The cells of third passage grew fastest, then slowed down and tended to stop in the fourth and fifth pas-sage, respectively. After several times of passage, the cells became larger and polymorphic in shape, some dendrite-like cells were observed. CONCLUSION：The method of twice-enzyme isolation may havest considerable cells in a short time with a high success rate. Morphology of DFCs is changed with the culture time and is multiple.

关 键 词：小鼠牙囊细胞 二次酶消化法 细胞培养 方法学 

分 类 号：R780.2[医药卫生—口腔医学]