摘要
目的 对口腔癌相关成纤维细胞 (CAFs)进行分离、培养、纯化并作初步鉴定。方法①用组织块培养法获得原代口腔CAFs和口腔黏膜正常成纤维细胞 (NFs) ,0 .2 5 %胰蛋白酶消化传代 ,联合采用机械刮除法和酶消化法纯化细胞 ;②通过形态学观察和多种蛋白的免疫组化染色 ,对口腔CAFs进行初步鉴定。结果 ①获得第 3代纯化口腔CAFs;②口腔CAFs呈长梭形 ,胞质突减少 ,细胞大小不一致 ,生长密集 ,排列紊乱。电镜下可见肌丝和电子致密斑等超微结构 ;③口腔CAFs的细胞角蛋白免疫组化染色呈阴性 ,波形蛋白、α 平滑肌动蛋白、基质金属蛋白酶 2染色呈阳性。结论CAFs与NFs相比 ,在形态结构、生长方式、蛋白表达等方面差异均有显著性 ,推测口腔肿瘤
Objective To separate, cultivate, purify and identify oral carcinoma associated fibroblasts (CAFs) preliminarily. Methods 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). Results 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, α smooth muscle actin and matrix metalloproteinases 2. Conclusions 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.
出处
《中华口腔医学杂志》
CAS
CSCD
北大核心
2004年第2期122-125,共4页
Chinese Journal of Stomatology
基金
四川省科委重点科技攻关基金资助项目(0 2kw 0 0 5 )
