Establishment and characterization of immortalized mouse ameloblast-like cell lines.
Orthod Craniofac Res
; 22 Suppl 1: 134-141, 2019 May.
Article
em En
| MEDLINE
| ID: mdl-31074144
ABSTRACT
OBJECTIVES:
Enamel organ epithelium (EOE) gives rise to the major epithelial-derived cell types of tooth including the ameloblasts. The formation of enamel, termed amelogenesis, occurs through the cytodifferentiation of ameloblasts, ultimately leading to apoptosis and necrosis of these cells with eruption. Therefore, studies regarding enamel matrix formation and bioengineering have been limited. In this study, we establish and characterize two mouse immortalized ameloblast-like cell lines using human papillomavirus 16 (HPV16) E6/E7 oncogenes for the first time. SETTING AND SAMPLE POPULATION Two mouse EOE dental cell lines (EOE-2M and EOE-3M). MATERIAL ANDMETHODS:
Isolated EOE primary cells were used to establish clonal cell lines and immortalized using the HPV16 E6/E7 gene platform. Two established cell lines were characterized by growth rate (Cell Proliferation Assay, MTS), gene (quantitative RT-PCR) and protein (immunocytochemistry) expression profiles, and mineralization potential (in situ alkaline phosphatase (ALP) histochemistry and Xylene Orange staining) in media supplemented with ascorbic acid and ß-glycerophosphate. Gene and protein expression analyses included specific enamel matrix and ameloblast cell markers Amel, Ambn, Enam, Amtn, ODAM, MMP20, Krt14 and DLX3.RESULTS:
Both cell lines were maintained in excess of 30 passages, with EOE-3M cells proliferating at a slightly higher rate. The cell lines expressed all tested enamel matrix markers and produced a mineralized ECM demonstrating an ameloblast-like profile.CONCLUSIONS:
Two mouse ameloblasts-like immortalized cell lines have been characterized that will be useful tool for studies regarding enamel bioengineering.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Dente
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Linhagem Celular
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Ameloblastos
Limite:
Animals
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Humans
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article