[Human oral carcinoma-associated fibroblasts: its cultivation, purification and identification].
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, alpha-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.
Zhou
Hong-mei
HM
Research Laboratory of Oral Mucosal Diseases, West China School of Stomatology, Sichuan University, Chengdu 610041, China. acomnet@163.net
Liu
Ying
Y
Hu
Tao
T
Li
Sheng-fu
SF
Gao
Qing-hong
QH
Li
Bing-qi
BQ
D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods*; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D009062:Mouth Neoplasms / Q000473:pathology*