Characterization of human dental pulp‐derived cell lines

Abstract Aim To establish and characterize different types of fibroblastic cell lines derived from dental pulp tissue. Methodology Human dental pulp tissue‐derived cells were transfected with SV40 large T antigen by Lipofectamine transfection method. Geneticin (G418)‐resistant cells were selected and different cell lines were established by a limiting dilution method. To characterize the lineages of cells, each clone was immunofluorescently stained by anti‐fibroblast, anti‐vimentin, anti‐collagen type I and type III antibodies. Total RNA was extracted from each clone and subjected to a differential display experiment. Results By transfecting SV40 large T antigen, nine different cell clones were obtained. All these cell clones were positively stained by anti‐fibroblast, anti‐vimentin, anti‐collagen type I and type III antibodies. With differential display experiment, eight different genes, the expression levels of these genes were varied amongst each cell clone, were detected. After sequencing and database search, one gene was revealed to be identical to T‐cell marker, Thy‐1. Thy‐1 expression in dental pulp tissue was confirmed by immunohistochemical staining. Conclusion Fibroblastic cell lines derived from human dental pulp tissue possessed different gene expression profiles suggesting the existence of subpopulations.