Document Detail


Establishment of human dental epithelial cell lines expressing ameloblastin and enamelin by transfection of hTERT and cdk4 cDNAs.
MedLine Citation:
PMID:  20923449     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: An in vitro cell culture system of dental epithelium is useful for the investigations of cellular differentiation and function of ameloblast in amelogenesis and of regenerative therapy in human tooth. However, there have been no immortalized human dental epithelial ameloblastic-lineage cell lines, which proliferate indefinitely and additionally produce enamel matrix proteins.
METHODS: We transfected two retroviral constructs of human telomerase reverse transcriptase (hTERT) cDNA and mouse cyclin-dependent kinase 4 (cdk4) cDNA into the primary ameloblastoma cells and isolated immortalized human dental epithelial cell lines of HAM1, HAM2 and HAM3. The three cell lines were examined by electron microscopy, assay of senescence-associated β-galactosidase activity, mRNA expression and immuno-reactivity of dental epithelial marker cell molecules and enamel matrix proteins.
RESULTS: They showed undifferentiated phenotypes in monolayer culture and did not have any β-galactosidase activity. The transcripts of dental epithelial cell markers of Msx2, Jagged1, Notch1, Sp3, Sp6, keratin 14 and keratin 18 were confirmed. In addition, mRNA and protein expression of ameloblastin and enamelin were also detected in three cell lines. All cells in the three cell lines were keratin 14- and 18-positive and some elongated cells were Jagged1-positive. Msx2-positive nuclei were noted in only HAM2 cells.
CONCLUSION: We established three cell lines by transfection of hTERT and cdk4 cDNAs, which were characterized as dental epithelial progenitor cells containing ameloblast-lineage cell phenotype.
Authors:
Setsuko Hatakeyama; Noriko Mizusawa; Reiko Tsutsumi; Katsuhiko Yoshimoto; Harumi Mizuki; Shigeru Yasumoto; Shigehiro Sato; Yasunori Takeda
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-10-04
Journal Detail:
Title:  Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology     Volume:  40     ISSN:  1600-0714     ISO Abbreviation:  J. Oral Pathol. Med.     Publication Date:  2011 Mar 
Date Detail:
Created Date:  2011-02-23     Completed Date:  2011-06-28     Revised Date:  2012-06-25    
Medline Journal Info:
Nlm Unique ID:  8911934     Medline TA:  J Oral Pathol Med     Country:  Denmark    
Other Details:
Languages:  eng     Pagination:  227-34     Citation Subset:  D; IM    
Copyright Information:
© 2010 John Wiley & Sons A/S.
Affiliation:
Department of Pathogenesis and Control of Oral Diseases, School of Dentistry, Iwate Medical University, Morioka, Japan. hsetsuko11@gmail.com
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MeSH Terms
Descriptor/Qualifier:
Ameloblastoma / pathology
Ameloblasts / cytology*
Animals
Calcium-Binding Proteins / analysis
Cell Culture Techniques
Cell Death
Cell Differentiation / physiology
Cell Line*
Cell Lineage
Cell Proliferation
Cell Separation
Cyclin-Dependent Kinase 4 / genetics
Dental Enamel Proteins / metabolism*
Epithelial Cells / cytology
Homeodomain Proteins / analysis
Humans
Intercellular Signaling Peptides and Proteins / analysis
Keratin-14 / analysis
Keratin-18 / analysis
Kruppel-Like Transcription Factors / analysis
Membrane Proteins / analysis
Mice
Mice, Inbred BALB C
Mice, Nude
RNA, Messenger / analysis
Receptor, Notch1 / analysis
Sp3 Transcription Factor / analysis
Telomerase / genetics
Transfection / methods*
beta-Galactosidase / analysis
Chemical
Reg. No./Substance:
0/AMBN protein, human; 0/Calcium-Binding Proteins; 0/Dental Enamel Proteins; 0/Homeodomain Proteins; 0/Intercellular Signaling Peptides and Proteins; 0/KRT14 protein, human; 0/KRT18 protein, human; 0/Keratin-14; 0/Keratin-18; 0/Kruppel-Like Transcription Factors; 0/MSX2 protein; 0/Membrane Proteins; 0/NOTCH1 protein, human; 0/RNA, Messenger; 0/Receptor, Notch1; 0/SP3 protein, human; 0/tuftelin; 134324-36-0/Serrate proteins; 148710-94-5/Sp3 Transcription Factor; EC 2.7.11.22/Cdk4 protein, mouse; EC 2.7.11.22/Cyclin-Dependent Kinase 4; EC 2.7.7.49/Telomerase; EC 3.2.1.23/beta-Galactosidase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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