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Establishment and characterization of a telomerase immortalized human gingival epithelial cell line
Author(s) -
MoffattJauregui C. E.,
Robinson B.,
Moya A. V.,
Brockman R. D.,
Roman A. V.,
Cash M. N.,
Culp D. J.,
Lamont R. J.
Publication year - 2013
Publication title -
journal of periodontal research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.31
H-Index - 83
eISSN - 1600-0765
pISSN - 0022-3484
DOI - 10.1111/jre.12059
Subject(s) - cytokeratin , telomerase , telomerase reverse transcriptase , biology , immortalised cell line , cell culture , senescence , epithelium , telomere , microbiology and biotechnology , porphyromonas gingivalis , keratinocyte , keratin , pathology , immunology , genetics , immunohistochemistry , medicine , dna , bacteria , gene
Background and Objective Gingival keratinocytes are used in model systems to investigate the interaction between periodontal bacteria and the epithelium in the initial stages of the periodontal disease process. Primary gingival epithelial cells ( GEC s) have a finite lifespan in culture before they enter senescence and cease to replicate, while epithelial cells immortalized with viral proteins can exhibit chromosomal rearrangements. The aim of this study was to generate a telomerase immortalized human gingival epithelial cell line and compare its in vitro behaviour to that of human GEC s. Material and Methods Human primary GECs were immortalized with a bmi1/ hTERT combination to prevent cell cycle triggers of senescence and telomere shortening. The resultant cell‐line, telomerase immortalized gingival keratinocytes ( TIGK s), were compared to GEC s for cell morphology, karyotype, growth and cytokeratin expression, and further characterized for replicative lifespan, expression of toll‐like receptors and invasion by P. gingivalis . Results TIGK s showed morphologies, karyotype, proliferation rates and expression of characteristic cytokeratin proteins comparable to GEC s. TIGK s underwent 36 passages without signs of senescence and expressed transcripts for toll‐like receptors 1–6, 8 and 9. A subpopulation of cells underwent stratification after extended time in culture. The cytokeratin profiles of TIGK monolayers were consistent with basal cells. When allowed to stratify, cytokeratin profiles of TIGK s were consistent with suprabasal cells of the junctional epithelium. Further, TIGK s were comparable to GEC s in previously reported levels and kinetics of invasion by wild‐type P. gingivalis and an invasion defective Δ serB mutant. Conclusion Results confirm bmi1/ hTERT immortalization of primary GECs generated a robust cell line with similar characteristics to the parental cell type. TIGK s represent a valuable model system for the study of oral bacteria interactions with host gingival cells.