Isolation and characterization of an immortalized oral keratinocyte cell line of mouse origin.
Isolation and characterization of an immortalized oral keratinocyte cell line of mouse origin.
复制标题
长老的口服角质形成细胞系的分离和表征小鼠起源。
DOI：
10.1016/j.archoralbio.2008.07.002
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发表时间：
2008-11
影响因子：
3
通讯作者：
Garrett-Sinha, Lee Ann
中科院分区：
文献类型：
--
作者：
Parikh, Neha;Nagarajan, Priyadharsini;Sei-Ichi, Matsui;Sinha, Satrajit;Garrett-Sinha, Lee Ann
目的：建立小鼠口腔上皮细胞系，进行分子和生化分析。从成年小鼠的口腔中分离上皮细胞，并在培养中建立一个自发永生的细胞系，称为永生口腔角质细胞（IMOK细胞）。然后对细胞的生长特性、分化潜力、核型、可转染性、对病毒感染的易感性和对siRNA的反应进行表征。IMOK细胞在含血清和无血清培养基中均表现出强劲的生长，至少可达到100倍的群体增长。IMOK细胞具有接近二倍体的核型，表达角质细胞标记蛋白，并且可以通过在培养基中添加高水平的钙来诱导分化。分化过程以形态学改变和诱导口腔上皮特异性分化标记蛋白如K4和K13为特征。瞬时转染分析表明，IMOK细胞具有高度可转染性，并且上皮细胞的几种启动子在这些细胞中具有活性。此外，在钙的作用下，分化特异性K4和Elf5启动子活性上调。最后，我们发现口腔角化细胞也可受逆转录病毒感染和基于siRNA的基因表达敲除。我们的研究首次建立了小鼠来源的永生化口腔角质细胞细胞系，该细胞系可以概括口腔上皮分化程序，因此可以作为口腔组织毒理学和分子分析的有用工具。
To establish an oral epithelial cell line of mouse origin for molecular and biochemical assays. Epithelial cells were isolated from the oral cavity of adult mice and established as a spontaneously immortalized cell line in culture, designated immortalized oral keratinocyte cells (IMOK cells). The cells were then characterized for growth characteristics, differentiation potential, karyotype, transfectability, susceptibility to viral infection and responses to siRNA. The IMOK cells exhibit robust growth in both serum-containing and serum-free medium for at least 100 population doublings. IMOK cells have a near diploid karyotype, express keratinocyte marker proteins and can be induced to undergo differentiation by addition of high levels of calcium to the medium. The differentiation process is characterized by morphological changes and by the induction of oral epithelium specific differentiation marker sproteins such as K4 and K13. Transient transfection analyses reveal that IMOK cells are highly transfectable and that several promoters of epithelial cells are active in these cells. Moreover, upon differentiation with calcium, there is an up-regulation of differentiation-specific K4 and Elf5 promoter activity. Finally, we show that the oral keratinocytes are also amenable to infection with retroviruses and to siRNA based knock-down of gene expression. Our study is the first to establish an immortalized oral keratinocyte cell line of murine origin that can recapitulate the oral epithelium differentiation program and thus could serve as a useful tool for toxicological and molecular analyses of the oral tissue.
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影响因子：
6.5
作者：
Romano, Rose-Anne;Birkaya, Barbara;Sinha, Satrajit
通讯作者：
Sinha, Satrajit
影响因子：
4.8
作者：
John, Shinu A.;Clements, James L.;Garrett-Sinha, Lee Ann
通讯作者：
Garrett-Sinha, Lee Ann
影响因子：
10.3
作者：
MARCELO, CL;GOLD, RC;FAIRLEY, JA
通讯作者：
FAIRLEY, JA
DOI：
10.1023/a:1016385109922
发表时间：
2001-01-01
期刊：
Methods in Cell Science
影响因子：
--
作者：
Hakkinen, Lari;Koivisto, Leeni;Larjava, Hannu
通讯作者：
Larjava, Hannu
影响因子：
14.8
作者：
Lichti, Ulrike;Anders, Joanna;Yuspa, Stuart H.
通讯作者：
Yuspa, Stuart H.