Staab Claudia A, Vondracek Martin, Custodio Hipolito, Johansson Katarina, Nilsson Jan Anders, Morgan Peter, Höög Jan-Olov, Cotgreave Ian, Grafström Roland C
Institute of Environmental Medicine, Karolinska Institute, Stockholm, Sweden.
Altern Lab Anim. 2004 Oct;32(4):401-5. doi: 10.1177/026119290403200412.
Normal oral keratinocytes (NOKs), and a Simian virus 40 T-antigen-immortalised oral keratinocyte line termed SVpgC2a, were cultured in an effort to model the human oral epithelium in vitro, including normal and dysplastic tissue. Monolayer and organotypic cultures of NOKs and SVpgC2a were successfully established in a standardised serum-free medium with high levels of amino acids, by using regular tissue culture plastic for monolayers and collagen gels containing oral fibroblasts as the base for generating tissue equivalents. NOKs express many characteristics of normal tissue, including those associated with terminal squamous differentiation. After > 150 passages, SVpgC2a cells retained an immortal, nontumourigenic phenotype that, relative to NOKs, was associated with aberrant morphology, enhanced proliferation, deficiency in terminal differentiation, proneness to apoptosis, and variably altered expression of structural epithelial markers. Transcript and protein profiling, as well as activity assays, demonstrated the expression of multiple xenobiotic-metabolising enzymes in SVpgC2a cells, some of which were higher in comparison to NOKs. A generally preserved, or even activated, ability for xenobiotic metabolism in long-term cultures of SVpgC2a cells indicated that this cell line could be useful in safety testing protocols--for example, in the development of consumer products in the oral health care field. However, SVpgC2a cells displayed some features reminiscent of a severe oral dysplasia, implying that this cell line could also to some extent serve as a model of a premalignant oral epithelium.
为了在体外模拟人类口腔上皮，包括正常组织和发育异常组织，培养了正常口腔角质形成细胞（NOKs）和一种名为SVpgC2a的猿猴病毒40 T抗原永生化口腔角质形成细胞系。通过使用常规组织培养塑料进行单层培养，并以含有口腔成纤维细胞的胶原凝胶作为生成组织等效物的基础，在含有高水平氨基酸的标准化无血清培养基中成功建立了NOKs和SVpgC2a的单层培养和器官型培养。NOKs表现出正常组织的许多特征，包括与终末鳞状分化相关的特征。经过150多代传代后，SVpgC2a细胞保留了一种永生的、非致瘤性表型，相对于NOKs，该表型与形态异常、增殖增强、终末分化缺陷、易凋亡以及结构上皮标志物的表达变化有关。转录组和蛋白质谱分析以及活性测定表明，SVpgC2a细胞中表达多种外源性代谢酶，其中一些酶的表达水平高于NOKs。SVpgC2a细胞长期培养中对外源性代谢的能力通常得以保留，甚至被激活，这表明该细胞系可用于安全测试方案，例如在口腔保健领域消费品的开发中。然而，SVpgC2a细胞表现出一些类似于严重口腔发育异常的特征，这意味着该细胞系在某种程度上也可作为癌前口腔上皮的模型。
