title: Characterization of human primary enamel organ epithelial cells in vitro.
abstract: 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.
authors.affiliation: University of California at San Francisco, 94143-0640, USA. pkdb@itsa.ucsf.edu
authors.forename: P K
authors.initials: PK
authors.lastname: DenBesten
authors.forename: D
authors.initials: D
authors.lastname: Machule
authors.forename: Y
authors.initials: Y
authors.lastname: Zhang
authors.forename: Q
authors.initials: Q
authors.lastname: Yan
authors.forename: W
authors.initials: W
authors.lastname: Li
mesh_terms: 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
chemical_list: 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