A simple and reliable technique for culturing of human oral keratinocytes and fibroblasts

Cultures of non-malignant oral cells are needed for many applications in the field of oral biology. We describe an efficient and rapid method for the culture of both keratinocytes and fibroblasts isolated from oral tissue, uvulas from patients undergoing reconstructive surgery and non-cancerous tissue from oral cancer patients. High cell yields with a high purity could be obtained. The technique is described in detail with respect to isolation of the cell types, the best way to propagate for over 5 passages and to measure cell proliferation in a colorimetric assay. Culture was successful in over 80% of cases for keratinocytes and over 63% for fibroblasts. Problems encountered were the risk of microbiological infection, the size of the specimen and the type of culture medium used. The potential applications of these primary cell cultures are mentioned.