小鼠成釉细胞的离体培养和纯化

Culture and purification of mouse ameloblasts in vitro

目的:对C57BL/6小鼠成釉细胞进行离体培养,探寻适合小鼠成釉细胞的体外培养方法。方法:取新生C57BL/6乳鼠,体视显微镜下分离其上下颌第一磨牙牙胚,收集组织后用胰蛋白酶消化成细胞悬液,接种于BME包被的培养皿中。在倒置显微镜下观察细胞形态及生长状况,差速消化法纯化细胞后,用免疫荧光法检测细胞角蛋白14和釉原蛋白(AMELX)的表达情况。结果:原代培养的小鼠成釉细胞生长良好,其中混有大量成纤维样细胞,经改良纯化后,细胞呈铺路石样成片生长,K14和AMELX表达阳性。结论:采用基底膜抽提物作为基质,酶消化法培养后经差速消化法纯化得到小鼠成釉细胞。

AIM： To culture mouse ameloblasts in vitro. METHODS： First molar tooth germs from new-born C57 BL /6 mice were obtained under stereoscopic microscope. The tooth germ tissues were digested with trypticase and cultured on BME coated dish. The cultured cells were purified by differential digestion and observed under inverted microscope.The purified cells were examined for the expression of cytokeratin 14 and amelogenin（ AMELX） by immunofluorescence.RESULTS： The primary cultured ameloblasts were mixed with fibroblast-like cells. However,the purified ameloblasts were cobblestone-shaped,growing well,K14 and AMELX positive. CONCLUSION： Mouse ameloblasts can be in vitro cultured by enzyme digestion method,and purified by differential digestion in BME-coated dishes.