Zhou Hong-mei, Liu Ying, Hu Tao, Li Sheng-fu, Gao Qing-hong, Li Bing-qi
Research Laboratory of Oral Mucosal Diseases, West China School of Stomatology, Sichuan University, Chengdu 610041, China.
Zhonghua Kou Qiang Yi Xue Za Zhi. 2004 Mar;39(2):122-5.
To separate, cultivate, purify and identify oral carcinoma-associated fibroblasts (CAFs) preliminarily.
The primary oral CAFs and normal fibrolasts (NFs) of oral mucosa were obtained by tissue culture. Then cells were dissociated by 0.25% trypsin and purified by curettage method and trypsinization. Morphological characteristics were observed under light microscope and electron microscope. The certain proteins were examined by immunohistochemistry (SP method).
The third passage purified oral CAFs were maintained. The characteristics of shape and growth of the oral CAFs changed significantly comparing to the NFs. Myofilament and electron dense patch were showed in the oral CAFs by electron microscope. The oral CAFs showed negative staining for cytokeratin, and positive staining for vimentin, alpha-smooth muscle actin and matrix metalloproteinases-2.
There are obvious differences of the morphological characteristics and expression of certain proteins between the CAFs and NFs. The microecology of the oral tumor-host interface might be one of the most important factors affecting the CAFs.
初步分离、培养、纯化及鉴定口腔癌相关成纤维细胞（CAFs）。
通过组织培养获取口腔原发性CAFs及口腔黏膜正常成纤维细胞（NFs）。然后用0.25%胰蛋白酶消化细胞，采用刮除法和胰蛋白酶消化法进行纯化。在光学显微镜和电子显微镜下观察细胞形态特征。采用免疫组织化学（SP法）检测某些蛋白质。
获得了第三代纯化的口腔CAFs。与NFs相比，口腔CAFs的形态和生长特性发生了显著变化。电子显微镜下口腔CAFs可见肌丝和电子致密斑。口腔CAFs细胞角蛋白染色呈阴性，波形蛋白、α-平滑肌肌动蛋白和基质金属蛋白酶-2染色呈阳性。
CAFs与NFs在形态特征和某些蛋白质表达方面存在明显差异。口腔肿瘤-宿主界面的微生态可能是影响CAFs的最重要因素之一。
