差速传代纯化大鼠牙囊细胞

Purification of rat dental follicle cells by differential passage

目的:建立一种简捷的分离培养和纯化大鼠牙囊细胞的方法。方法:分离出生后6dSD大鼠上下颌第一和第二磨牙完整牙胚,剥离牙囊和成釉器,剪碎后酶消化并混合培养,再利用多次差速传代纯化牙囊细胞。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4代可获得纯化的牙囊细胞。倒置显微镜下观察牙囊细胞呈长梭形或三角形,免疫组化染色抗波形丝蛋白阳性,抗角蛋白阴性。结论:利用多次差速传代可从混合培养的原代细胞中获得纯化的牙囊细胞。

Objective：To develop a simple culture and purifying method for rat dental follicle cells. Methods：The upper and lower first and second intact molar germs of SD rat were separated. Then, dental follicle and enamel organ were stripped together, minced into little pieces, digested with collagenase and cultured. Dental follicular cells were purified by differential passage and indentified by immunohistochemical staining of vimentin and cytokeratin. Results：The primary cells were mixed, consisting of dental follicle cells and enamel organ cells. After differential passage, the cells of fourth passage became purified dental follicle cells. Purified dental follicle cells were elongated spindle or triangle in shape, positive for vimentin and negative for cytokeratin. Conclusion： Dental follicle cells can be purified by several differential passages from the mixed primarily cultured cells.