大鼠成釉器细胞的分离纯化

目的：寻找一种大鼠成釉器细胞分离纯化的简捷方法。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，酶消化法原代混合培养。采用多次差别消化法去除成纤维样细胞，纯化上皮样细胞，并进行抗角蛋白染色和RT—PCR检测釉原蛋白、成釉蛋白mRNA表达。结果：原代细胞为混杂细胞，经2～3次差别消化可完全去除成纤维样细胞。上皮样细胞呈多角形，铺路石样生长，抗角蛋白染色阳性，表达釉原蛋白、成釉蛋白mRNA。结论：本研究通过多次差别消化获得了纯化的大鼠成釉器细胞。

Isolation and purification of rat enamel organ cells

AIM: To explore a simple approach to the isolation and purification of rat enamel organ cells.METHODS:The first and second molar germs were isolated from SD rat jaws.Dental follicle and enamel organ tissue were stripped from the germs and digested by enzyme for cell culture.Fibroblast-like cells were removed by differential digestion.The purified epithelial-like cells were immunostained with anti-keratin antibody.The expression of amelogenin and ameloblastin mRNA in epithelial-like cells were detected by RT-PCR.RESULTS:The primary culture was heterogeneous and the fibroblast-like cells could be completely removed after 2 to 3 times of differential digestion.The purified epithelial-like cells were polygonal and cobblestone-shape,immunopositive for keratin,and expressed amelogenin and ameloblastin mRNA.CONCLUSION: Purified enamel organ cells can be obtained by repeated differential digestion.