Characterization of human dental pulp-derived cell lines.
OBJECTIVE
To establish and characterize different types of fibroblastic cell lines derived from dental pulp tissue.


METHODS
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.


CONCLUSIONS
Fibroblastic cell lines derived from human dental pulp tissue possessed different gene expression profiles suggesting the existence of subpopulations.
Suguro
H
H
Department of Endodontics, Dental Research Center, Nihon University School of Dentistry, Nihon Univeristy, Chiyoda-ku, Tokyo, Japan.
Asano
M
M
Kaneko
Y
Y
Omagari
D
D
Ogiso
B
B
Moro
I
I
Komiyama
K
K
D000328:Adult; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics; D002461:Cell Line, Transformed; D002999:Clone Cells; D003782:Dental Pulp / Q000166:cytology* / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D007124:Immunoenzyme Techniques; D017422:Sequence Analysis, DNA; D018800:Thy-1 Antigens / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
D000952:Antigens, Polyomavirus Transforming; D018800:Thy-1 Antigens