Characterization of human primary enamel organ epithelial cells in vitro.
Tooth enamel is formed by ameloblasts, which are derived from the epithelial cells of the enamel organ.


OBJECTIVE
The purpose of this study was to grow human ameloblast-like epithelial cells in culture.


METHODS
Human fetal tooth organs were isolated, and the cells were separated by digestion in collagenase/dispase. The cells were cultured in KGM-2 media with and without serum and at different calcium concentrations. The expression of enamel matrix proteins was analyzed by RT-PCR and cytokeratin 14 was detected by immunohistochemistry. The cells were further characterized by osteogenesis/odontogenesis-related DNA array.


RESULTS
Cells isolated from the tooth organs grown in KGM-2 media containing 2-10% serum, were mixture of cobblestone and spindle shaped cells. Culturing these cells in KGM-2 with 0.05 mM calcium was selective for cobblestone ameloblasts-like cells (CAB), which were immunopositive for cytokeratin 14. Amelogenin, ameloblastin, enamelin, MMP-20 and KLK-4 were detected in CAB cells by RT-PCR. Osteogenesis SuperArray analyses could not detect the presence of typical molecules related to mesenchymal odontoblast or osteoblast lineage cells in these cultures.


CONCLUSIONS
These studies showed that cobblestone-shaped ameloblast-like cells are selected from the tooth organ cells, by culture in KGM-2 media with 0.05 mM calcium.
DenBesten
P K
PK
University of California at San Francisco, 94143-0640, USA. pkdb@itsa.ucsf.edu
Machule
D
D
Zhang
Y
Y
Yan
Q
Q
Li
W
W
D000565:Ameloblasts / Q000502:physiology*; D053523:Amelogenin; D002102:Cadaver; D002118:Calcium / Q000032:analysis; D002454:Cell Differentiation / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D003746:Dental Enamel Proteins / Q000032:analysis; D004658:Enamel Organ / Q000166:cytology*; D004847:Epithelial Cells / Q000502:physiology; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D007610:Kallikreins / Q000032:analysis; D007633:Keratins / Q000032:analysis; D053519:Matrix Metalloproteinase 20; D020782:Matrix Metalloproteinases / Q000032:analysis; D008565:Membrane Proteins / Q000032:analysis; D009805:Odontogenesis / Q000502:physiology; D010641:Phenotype; D014083:Tooth Germ / Q000166:cytology* / Q000196:embryology
C490889:AMBN protein, human; D053523:Amelogenin; D003746:Dental Enamel Proteins; D008565:Membrane Proteins; C038456:tuftelin; D007633:Keratins; D007610:Kallikreins; C120801:kallikrein 4; C507111:MMP20 protein, human; D053519:Matrix Metalloproteinase 20; D020782:Matrix Metalloproteinases; D002118:Calcium