Bao, Kai ORCID: 0000-0002-1183-5607, Akguel, Baki and Bostanci, Nagihan (2014). Establishment and Characterization of Immortalized Gingival Epithelial and Fibroblastic Cell Lines for the Development of Organotypic Cultures. Cells Tissues Organs, 199 (4). S. 228 - 238. BASEL: KARGER. ISSN 1422-6421

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Abstract

In vitro studies using 3D co-cultures of gingival cells can resemble their in vivo counterparts much better than 2D models that typically only utilize monolayer cultures with short-living primary cells. However, the use of 3D gingival models is still limited through lack of appropriate cell lines. We aimed to establish immortalized cell line models of primary human gingival epithelium keratinocytes (HGEK) and gingival fibroblasts (GFB). Immortalized cell lines (HGEK-16 and GFB-16) were induced by E6 and E7 oncoproteins of human papillomavirus. In addition, 3D multilayered organotypic cultures were formed by embedding GFB-16 cells within a collagen (Col) matrix and seeding of HGEK-16 cells on the upper surfaces. Cell growth was analyzed in both immortalized cell lines and their parental primary cells. The expression levels of cell type-specific markers, i.e. cytokeratin (CK) 10, CK13, CK16, CK18, CK19 for HGEK-16 and Col I and Col II for GFB-16, were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). Expansion of the primary cultures was impeded at early passages, while the transformed immortalized cell lines could be expanded for more than 30 passages. In 3D cultures, immortalized HGEK formed a multilayer of epithelial cells. qRT-PCR showed that cell-specific marker expression in the 3D cultures was qualitatively and quantitatively closer to that in human gingival tissue than to monolayer cultures. These results indicate that immortalized gingival fibroblastic and epithelial cell lines can successfully form organotypic multilayered cultures and, therefore, may be useful tools for studying gingival tissue in vitro. (C) 2014 S. Karger AG, Basel

Item Type: Journal Article
Creators:
CreatorsEmailORCIDORCID Put Code
Bao, KaiUNSPECIFIEDorcid.org/0000-0002-1183-5607UNSPECIFIED
Akguel, BakiUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Bostanci, NagihanUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
URN: urn:nbn:de:hbz:38-449047
DOI: 10.1159/000363694
Journal or Publication Title: Cells Tissues Organs
Volume: 199
Number: 4
Page Range: S. 228 - 238
Date: 2014
Publisher: KARGER
Place of Publication: BASEL
ISSN: 1422-6421
Language: English
Faculty: Unspecified
Divisions: Unspecified
Subjects: no entry
Uncontrolled Keywords:
KeywordsLanguage
PERIODONTAL-LIGAMENT CELLS; VITRO SUBGINGIVAL BIOFILMS; IN-VITRO; CONNECTIVE-TISSUE; EXPRESSION; DIFFERENTIATION; KERATINOCYTES; RECEPTOR; DISEASE; MUCOSAMultiple languages
Anatomy & Morphology; Cell Biology; Developmental BiologyMultiple languages
URI: http://kups.ub.uni-koeln.de/id/eprint/44904

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