Separation, cultivation and biological characteristics of oral carcinoma-associated fibroblasts

Oral Dis. 2006 Jul;12(4):375-80. doi: 10.1111/j.1601-0825.2005.01207.x.

Abstract

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.

MeSH terms

  • Actins / analysis
  • Carcinoma, Squamous Cell / chemistry
  • Carcinoma, Squamous Cell / pathology*
  • Case-Control Studies
  • Cell Culture Techniques
  • Cell Proliferation
  • Cell Separation
  • Cell Survival
  • Cell Transformation, Neoplastic
  • Female
  • Fibroblasts / chemistry
  • Fibroblasts / pathology*
  • Humans
  • Karyotyping
  • Male
  • Matrix Metalloproteinase 2 / analysis
  • Mitotic Index
  • Mouth Mucosa / chemistry
  • Mouth Mucosa / cytology
  • Mouth Mucosa / pathology
  • Mouth Neoplasms / chemistry
  • Mouth Neoplasms / pathology*
  • Tumor Cells, Cultured
  • Vimentin / analysis

Substances

  • Actins
  • Vimentin
  • Matrix Metalloproteinase 2