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ID 112952
Title Alternative
in vitro 細胞培養系によるアメロジェネシス不全メカニズムの解読
IN VITRO MODELING OF AMELOGENESIS
Author
Arinawati, Dian Yosi Tokushima University
Hagita, Hiroko Tokushima University
Keywords
Amelogenesis imperfecta
Cell-cell interaction
Cell-matrix interaction
Dental epithelial cell
Genetic disease
In vitro amelogenesis imperfecta model
Phenotypic screening
Sp6
Content Type
Thesis or Dissertation
Description
The conventional two-dimensional (2D) in vitro culture system is frequently used to analyze the gene expression with or without extracellular signals. However, the cells derived from primary culture and cell lines frequently deviate the gene expression profile compared to the corresponding in vivo samples, which sometimes misleads the actual gene regulation in vivo. To overcome this gap, we developed the comparative 2D and 3D in vitro culture systems and applied them to the genetic study of amelogenesis imperfecta (AI) as a model. Recently, we found specificity protein 6 (Sp6) mutation in an autosomal-recessive AI rat that was previously named AMI. We constructed 3D structure of ARE-B30 cells (AMI-derived rat dental epithelial cells) or G5 (control wild type cells) combined with RPC-C2A cells (rat pulp cell line) separated by the collagen membrane, while in 2D structure, ARE-B30 or G5 was cultured with or without the collagen membrane. Comparative analysis of amelogenesis-related gene expression in ARE-B30 and G5 using our 2D and 3D in vitro systems revealed distinct expression profiles, showing the causative outcomes. Bone morphogenetic protein 2 and follistatin were reciprocally expressed in G5, but not in ARE-B30 cells. All-or-none expression of amelotin, kallikrein-related peptidase 4, and nerve growth factor receptor was observed in both cell types. In conclusion, our in vitro culture systems detected the phenotypical differences in the expression of the stage-specific amelogenesis-related genes. Parallel analysis with 2D and 3D culture systems may provide a platform to understand the molecular basis for defective amelogenesis caused by Sp6 mutation.
Journal Title
Journal of Bioscience and Bioengineering
ISSN
13891723
NCID
AA11644703
AA11307678
Publisher
The Society for Biotechnology, Japan|Elsevier
Volume
125
Issue
4
Start Page
479
End Page
489
Published Date
2018-02-01
Remark
内容要旨・審査要旨・論文本文の公開
本論文は,著者Dian Yosi Arinawatiの学位論文として提出され,学位審査・授与の対象となっている。
Rights
©2017. This manuscript version is made available under the CC-BY-NC-ND 4.0 license.( http://creativecommons.org/licenses/by-nc-nd/4.0 )
EDB ID
DOI (Published Version)
URL ( Publisher's Version )
FullText File
language
eng
TextVersion
ETD
MEXT report number
甲第3205号
Diploma Number
甲口第443号
Granted Date
2018-09-13
Degree Name
Doctor of Dental Science
Grantor
Tokushima University
departments
Oral Sciences