Development and characterization of an SV40 immortalized porcine ameloblast-like cell line.
Enamel is secreted as a protein matrix by the ameloblasts. These same cells then control the maturation of the enamel matrix, secreting proteinases that hydrolyze proteins as mineralization progresses, until mature enamel containing less than 1% protein by weight remains. Further understanding of the factors that control ameloblast function and differentiation requires an in vitro cell culture system. In this study, we report immortalization of enamel organ epithelial cells and the selection of a cell line with characteristics of ameloblasts. Porcine enamel organ cells were dissected from unerupted porcine molars, cultured in serum-free medium, and passaged twice. These cells were transfected with an origin-of-replication defective SV40 plasmid by calcium phosphate precipitation, and a cell line with mRNA expression characteristic of ameloblasts was cloned. This cell line (PABSo-E) expressed mRNA for amelogenin, matrix metalloproteinase-20 (enamelysin), and enamel matrix serine proteinase 1 (EMSP1), but not ameloblastin. PABSo-E cells have been passaged more than 55 times, while continuing to maintain characteristics of ameloblasts. These cells will be useful for future studies of ameloblast function.
DenBesten
P K
PK
Department of Growth and Development, University of California San Francisco, 94143-0640, USA. PKDB@itsa.ucsf.edu
Gao
C
C
Li
W
W
Mathews
C H
CH
Gruenert
D C
DC
D000565:Ameloblasts / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism / Q000502:physiology; D053523:Amelogenin; D000818:Animals; D002130:Calcium Phosphates; D002454:Cell Differentiation; D002461:Cell Line, Transformed; D011232:Chemical Precipitation; D002999:Clone Cells / Q000166:cytology; D016895:Culture Media, Serum-Free; D003743:Dental Enamel / Q000378:metabolism; D003746:Dental Enamel Proteins / Q000235:genetics / Q000378:metabolism; D004658:Enamel Organ / Q000166:cytology; D010450:Endopeptidases / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology; D007610:Kallikreins; D053519:Matrix Metalloproteinase 20; D020782:Matrix Metalloproteinases; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D012333:RNA, Messenger / Q000235:genetics; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism; D013539:Simian virus 40 / Q000235:genetics; D013552:Swine; D014074:Tooth Calcification / Q000502:physiology; D014083:Tooth Germ / Q000166:cytology; D014162:Transfection
D053523:Amelogenin; D002130:Calcium Phosphates; D016895:Culture Media, Serum-Free; D003746:Dental Enamel Proteins; D012333:RNA, Messenger; C485828:alpha-tricalcium phosphate; C490891:ameloblastin protein, Sus scrofa; C485830:tetracalcium phosphate; C485838:calcium phosphate, monobasic, anhydrous; C020243:calcium phosphate; D010450:Endopeptidases; D007610:Kallikreins; D012697:Serine Endopeptidases; C120801:kallikrein 4; D053519:Matrix Metalloproteinase 20; D020782:Matrix Metalloproteinases; D008666:Metalloendopeptidases; C485829:calcium phosphate, dibasic, anhydrous