Human oral carcinoma-associated fibroblasts: its cultivation, purification and identification.
- Author:
Hong-mei ZHOU
1
;
Ying LIU
;
Tao HU
;
Sheng-fu LI
;
Qing-hong GAO
;
Bing-qi LI
Author Information
- Publication Type:Journal Article
- MeSH: Cell Culture Techniques; methods; Cell Separation; methods; Fibroblasts; cytology; Humans; Mouth Neoplasms; pathology
- From: Chinese Journal of Stomatology 2004;39(2):122-125
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo separate, cultivate, purify and identify oral carcinoma-associated fibroblasts (CAFs) preliminarily.
METHODSThe 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).
RESULTSThe 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.
CONCLUSIONSThere 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.