Separation, cultivation and biological characteristics of oral carcinoma-associated fibroblasts
Separation, cultivation and biological characteristics of oral carcinoma-associated fibroblasts
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口腔癌相关成纤维细胞的分离、培养及生物学特性
DOI：
10.1111/j.1601-0825.2005.01207.x
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发表时间：
2006-07-01
期刊：
影响因子：
3.8
通讯作者：
Zhou, H. M.
中科院分区：
文献类型：
--
作者：
Liu, Y.;Hu, T.;Zhou, H. M.
目的：肿瘤相关成纤维细胞(CAF)被认为对多种实体肿瘤的发生和发展起调控作用。本研究的目的是对口腔CAF进行分离、培养、鉴定，并对其生物学特性进行研究。材料与方法：通过组织培养获得原代口腔CAF和正常舌成纤维细胞。0.25%胰酶分离细胞，刮除结合胰酶纯化。通过细胞形态观察和免疫组织化学染色对细胞进行鉴定。结果：成功地获得了第3代纯化的口腔CAF和NFSF。CAF的形态特征与非CAF相比发生了显著变化。CAF表达波形蛋白、α-平滑肌肌动蛋白和基质金属蛋白酶-2。CAF的增殖能力和有丝分裂能力均明显高于对照组(P<；0.05)。结论：口腔CAF与非口腔CAF的生物学特性存在明显差异。这一结果为进一步研究CAF在口腔癌发生发展中的作用提供了实验基础。
OBJECTIVES: Carcinoma-associated fibroblasts (CAFs) have been suggested to regulate the initiation and progression of many types of solid tumors. The aim of the study was to separate, cultivate, identify oral CAFs, and to investigate their biological characteristics.MATERIALS AND METHODS: The primary CAFs and normal fibroblasts (NFs) of the tongue were obtained by tissue culture. Then cells were dissociated by 0.25% trypsin and purified by curettage method combining with trypsinization. The cells were verified according to morphological observation and immunohistochemical staining of certain proteins. Multiple proliferation indexes and karyotype of the cells were assayed.RESULTS: Third passage purified oral CAFs and NFs were attained successfully. The morphological characteristics of the CAFs changed significantly comparing to the NFs. The CAFs showed positive staining for vimentin, alpha-smooth muscle actin and matrix metalloproteinases-2. The proliferation and mitosis ability of the CAFs were significantly increased compared with the NFs (P < 0.05). No karyotypic abnormalities were found in the CAFs.CONCLUSIONS: There were obvious differences in the biological characteristics between oral CAFs and NFs. The results may provide us an experimental foundation for further studies on the roles of CAFs in the initiation and progression of oral cancer.