Characterization of a system of mineralized-tissue formation by rat dental pulp cells in culture.
Pulp tissue was obtained from maxillary incisors of young adult male Wistar rats, minced and digested with 0.5% trypsin and 0.02% EGTA at 37 degrees C for 30 min. Dissociated cells were cultured with or without 10 nM dexamethasone using Eagle's minimal essential medium supplemented with 10% fetal bovine serum and 50 micrograms/ml ascorbic acid. Confluent cells were subcultured at 7 days and the medium further supplemented with beta-glycerophosphate (beta-GP). Dexamethasone in primary culture and/or secondary culture enhanced the formation of mineralized tissue while > 5 mM beta-GP was necessary for mineralization to occur. Biochemical analysis of the radiolabelled medium revealed that these cells produced type I, type I trimer and type III collagens. Analysis of [32PO4]-labelled medium, using DEAE-Sephacel ion-exchange chromatography and sodium dodecylsulphate-polyacrylamide gel electrophoresis, showed that these cells produced phosphophoryn-like protein. These results indicate that some of the rat dental pulp cells in culture express an odontoblast-like phenotype.
Kasugai
S
S
Department of Pharmacology, Faculty of Dentistry, Tokyo Medical and Dental University, Japan.
Shibata
S
S
Suzuki
S
S
Susami
T
T
Ogura
H
H
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D002848:Chromatography, DEAE-Cellulose; D003094:Collagen / Q000096:biosynthesis; D003782:Dental Pulp / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D003784:Dental Pulp Calcification / Q000139:chemically induced / Q000378:metabolism*; D003907:Dexamethasone / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005994:Glycerophosphates / Q000494:pharmacology; D008297:Male; D008437:Maxilla; D008858:Microscopy, Phase-Contrast; D009804:Odontoblasts; D010750:Phosphoproteins / Q000096:biosynthesis; D051381:Rats; D017208:Rats, Wistar
D005994:Glycerophosphates; D010750:Phosphoproteins; C017630:phosphophoryn; D003907:Dexamethasone; D003094:Collagen; D002118:Calcium; C031463:beta-glycerophosphoric acid