富含干细胞的鼠切牙颈环上皮细胞的分离和原代培养

Isolation and primary culture of cervical-loop epithelial cell from rat incisor

目的：获取较纯的鼠切牙颈环上皮细胞。方法：分离出生2d的Wistar大乳鼠下颌切牙完整的牙胚，机械剥离颈环组织，酶消化法与组织块法结合培养原代混合细胞。采用Ⅳ型胶原包被培养法纯化上皮干细胞。结果：原代培养的细胞为混杂细胞，经2—3次纯化可完全去除纤维样细胞。颈环上皮细胞呈多角形，克隆状生长。结论：Ⅳ型胶原包被培养法可以获取纯化的鼠颈环上皮细胞。

AIM： To obtain purified cervical - loop epithelial cells in lower incisor of wistar rat . METHODS ： The lower incisor tooth germs were dissected from the mandibles of two - day - old wistar rats. Both the labial and lingual epitheliums were mechanically stripped from the incisor tooth germs, leaving only cervical - loop tissues. The cervical- loop tissues were cultured by enzymolysis and tissue cultutre methods and purified by coating collagen type IV. The purified cells were then identified by immunohistochemistry staining. RESULTS： The primary culture was a mixture of different cells. After two - or three - time purification, the fibroblast - like cells could be fully removed. The cervical - loop epithelial cells were polygonal and able to form clones. The cerivical - loop epithelial cells exhibited positive expression for integrin - β1 and monoclonal antibody CK in immunohistochemistry staining. CONCLUSION ： The purified cervical - loop epithelial cells in rat lower incisors can be obtained by coating collagen type Ⅳ.