Establishment and characterization of a telomerase immortalized human gingival epithelial cell line.
Establishment and characterization of a telomerase immortalized human gingival epithelial cell line.
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DOI：
10.1111/jre.12059
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发表时间：
2013-12
影响因子：
3.5
通讯作者：
Lamont RJ
中科院分区：
文献类型：
--
作者：
Moffatt-Jauregui CE;Robinson B;de Moya AV;Brockman RD;Roman AV;Cash MN;Culp DJ;Lamont RJ
在模型系统中使用牙龈角质形成细胞来研究在牙周病过程的初始阶段牙周细菌和上皮之间的相互作用。原代牙龈上皮细胞（GEC）在进入衰老并停止复制之前在培养物中具有有限的寿命，而用病毒蛋白永生化的上皮细胞可表现出染色体重排。本研究的目的是产生端粒酶永生化的人牙龈上皮细胞系，并比较其在体外的行为，人类GECs。人原代牙龈上皮细胞永生化与bmi 1/hTERT组合，以防止细胞周期触发衰老和端粒缩短。将所得细胞系端粒酶永生化牙龈角质形成细胞（TIGKs）与GEC的细胞形态、核型、生长和细胞角蛋白表达进行比较，并进一步表征复制寿命、toll样受体（TLR）的表达和牙龈卟啉单胞菌的侵袭。TIGKs的形态、核型、增殖率和特征性细胞角蛋白的表达与GECs相当。TIGKs经历了36次传代，没有衰老迹象，并表达TLR 1-6、8和9的转录物。细胞亚群在延长的培养时间后经历分层。TIGK单层的细胞角蛋白谱与基底细胞一致。当允许分层时，TIGKs的细胞角蛋白谱与交界上皮的基底上细胞一致。此外，TIGKs在先前报道的野生型牙龈卟啉单胞菌和侵袭缺陷型ΔserB突变体的侵袭水平和动力学方面与GEC相当。结果证实，原代牙龈上皮细胞的bmi 1/hTERT永生化产生了一个强大的细胞系，具有类似的特征，以父母的细胞类型。TIGKs是研究口腔细菌与宿主牙龈细胞相互作用的有价值的模型系统。
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 (GECs) 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 behavior to that of human GECs. Human primary gingival epithelial cells 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 (TIGKs), were compared to GECs for cell morphology, karyotype, growth and cytokeratin expression, and further characterized for replicative lifespan, expression of toll-like receptors (TLRs) and invasion by P. gingivalis. TIGKs showed morphologies, karyotype, proliferation rates and expression of characteristic cytokeratin proteins comparable to GECs. TIGKs underwent 36 passages without signs of senescence and expressed transcripts for TLRs 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 TIGKs were consistent with suprabasal cells of the junctional epithelium. Further, TIGKs were comparable to GECs in previously reported levels and kinetics of invasion by wild type Porphyromonas gingivalis and an invasion defective ΔserB mutant. Results confirm bmi1/hTERT-immortalization of primary gingival epithelial cells generated a robust cell-line with similar characteristics to the parental cell type. TIGKs represent a valuable model system for the study of oral bacteria interactions with host gingival cells.
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影响因子：
3.7
作者：
Dickinson BC;Moffatt CE;Hagerty D;Whitmore SE;Brown TA;Graves DT;Lamont RJ
通讯作者：
Lamont RJ
影响因子：
5.2
作者：
Dabelsteen, Sally;Hercule, Paula;Barron, Patricia;Rice, Meghan;Dorsainville, Gregory;Rheinwald, James G.
通讯作者：
Rheinwald, James G.
影响因子：
3.4
作者：
Handfield, M;Mans, JJ;Lamont, RJ
通讯作者：
Lamont, RJ
影响因子：
3.2
作者：
Das, Biswadip;Cash, Melanie N.;Culp, David J.
通讯作者：
Culp, David J.
影响因子：
3.4
作者：
Mao, Song;Park, Yoonsuk;Lamont, Richard J.
通讯作者：
Lamont, Richard J.