双向差速法纯化大鼠牙囊细胞  

Rat dental follicle cells purification by bidirectional differential method

作　　者：王丽萍[1] 李伯琦[1] 铁晓敏 王琪[1] 刘奕杉[1] 

机构地区：[1]新疆医科大学第一附属医院儿童口腔科-口腔预防科,新疆乌鲁木齐830054

出　　处：《口腔疾病防治》2016年第3期142-145,共4页Journal of Prevention and Treatment for Stomatological Diseases

基　　金：国家自然科学基金(81560178)

摘　　要：目的利用牙囊细胞和成釉细胞贴壁速度及酶消化分离速度不同的特点,建立一种简便、快速纯化大鼠牙囊细胞的方法。方法取新生5~6 d SD大鼠下颌第一磨牙牙胚,在体视显微镜下剥离牙囊及成釉器,剪碎后酶消化并混合培养,再利用差速贴壁法和差速传代法纯化牙囊细胞。结果原代细胞为牙囊细胞和成釉器细胞混合生长,差速传代培养到第2~3代可获得纯化的牙囊细胞。倒置显微镜下观察牙囊细胞呈梭形或三角形,免疫组织化学染色显示抗波形丝蛋白阳性,抗角蛋白阴性。结论双向差速法是一种高效、简便的纯化牙囊细胞的方法。Objective To develop a simple and fast purifingmethod of dental follicle cell by the character of different velocity of attachment of detachment between dental follicle cells and enamel organ cells. Methods The lower first intact molar germs of SD rats with 5-6 days old were selected and separated. The dental follicle and enamel organ were stipped together under the stereo microscope. After minced into little pieces and degested with trypsin and cultured, the dental follicle cells were purified by diffierential adherence and differential passage. The final purified cells were identified by immunocytochemistry. Results The primay cells were mixed, consisting of dental follice cells and enamel organ cells.After differential adherence and passage, the cells of 2-3th passage became purified dental follicle cells. Purified dental follicle cells were elongate spindle or triangle in shape, positive for vimentin and negative for cytokeratin. Conclusion Bidirectional differential method is an efficient and simple method for the purification of dental follicle cells.

关 键 词：牙囊细胞 纯化 差速贴壁 差速传代 免疫组织化学 

分 类 号：Q2[生物学—细胞生物学]