Banerjee N Sanjib, Chow Louise T, Broker Thomas R
Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, USA.
Methods Mol Med. 2005;119:187-202. doi: 10.1385/1-59259-982-6:187.
The productive phase of human papillomavirus (HPV) infection is dependent on squamous differentiation of epithelial keratinocytes. Organotypic culture systems of primary human keratinocytes (PHKs) or immortalized keratinocytes that contain HPV genomes were developed to recapitulate this permissive environment. A complementary approach to determine the functions of individual HPV genes and to examine the virus-host interactions is to introduce the gene, alone or in combination, into keratinocytes that are then grown in organotypic cultures. The success of the latter approach depends on the methodology of retrovirus-mediated gene transfer, which can transduce the viral gene or genes into an entire population of PHKs. In this chapter, we describe the strategies and methods of retrovirus-mediated gene transfer into keratinocytes grown into organotypic cultures.
人乳头瘤病毒（HPV）感染的增殖阶段依赖于上皮角质形成细胞的鳞状分化。已开发出包含HPV基因组的原代人角质形成细胞（PHK）或永生化角质形成细胞的器官型培养系统，以重现这种允许性环境。确定单个HPV基因功能并检查病毒与宿主相互作用的一种补充方法是将该基因单独或组合引入角质形成细胞，然后在器官型培养物中培养。后一种方法的成功取决于逆转录病毒介导的基因转移方法，该方法可将一个或多个病毒基因转导到整个PHK群体中。在本章中，我们描述了逆转录病毒介导的基因转移到生长在器官型培养物中的角质形成细胞的策略和方法。
