Human Dental Papilla Cells In Culture And The Study Of Their Biological Characteristics

The dental papilla cells represent a unique embryonic mesenchymal cell population which plays a very important regulatory role during tooth development. They have attracted considerable attention because these cells contain information on the size and shape of the tooth. The ability to maintain isolated cells in culture provides a powerful tool to study many aspects of cell function, especially odontoblast differentiation.To the best of our knowledge, there have been several reports on the characterization of mouse dental papilla cells cultured in vitro, but few reports on the human dental papilla cells. The purpose of this thesis is to establish a stable dental papilla cell line of human embryonic origin and invest the effects of 1,25-dihydroxyvitamin D₃[1,25(OH)₂D₃], a strong differentiation inducer, epidermal growth factor(EGF), insulin and transforming growth factor- β (TGF- β) on human dental papilla cells in vitro. The thesis includes three parts. 1.The establishment of a model of Culturing human dental papilla cells. 2. Effects of 1,25(OH)₂D₃, EGF, Insulin and TGF- β on the biological characters of human dental papilla cells in vitro. 3. An in situ hybridization study of Calbindin-D28k(D28k) in human embryonic incisor tooth germ and dental papilla cells in vitro. The results showed as below:The mandible of 4 to 6-month-old human embryo was obtained via legal abortion induced with non-medication method. The dental papilla mesenchyme were manually dissociated and chopped to around 1 mm³ and cultured in DMEM or RPMI 1640 supplemented with 15% fetal calf serum ,100U/ml penicillin and streptomycin separately. Cultures were maintained at 37℃ in a humidified atmosphere of 5% CO₂, and 95% air.