大鼠根尖蕾细胞的分离培养

Separation and culture of rats' apical bud cells

目的研究大鼠切牙根尖蕾细胞的分离、培养和纯化的方法。方法分离出生后3dSD大鼠下颌切牙胚,切取根尖唇侧部分组织,酶消化法原代培养,利用差速传代纯化上皮细胞。用角蛋白14和波形丝蛋白染色确定细胞来源。结果原代培养的细胞为上皮和间充质混合细胞,经2～3次差速传代后可获得纯化的根间蕾上皮细胞。免疫组化染色角蛋白14阳性,波形丝蛋白阴性。结论利用酶消化法和差速传代法培养获得了纯化的根尖蕾细胞。

Objective To develop a culture method of apical bud cells. Methods We separated the lower incisor germs of post natal 3- day SD rats. We cut the labial apical part of these germs and cultured the mixed cells by the digestive method. Then, the epithelial cells were purified by several differential passages and identified by immunohistochemical staining of cytokeratinl4 and vimentin. Results The primary cells were mixed by epithelial and mesenchymal cells. After 2 - 3 differential passages, purified epithelial cells were harvested, which were positively stained for cytokeratinl4 and negatively for vimentin. Conclusions Apical bud cells can be cultured by the digestive method and purified by differential passages.