McMaster Immunology Research Center, McMaster University, Department of Pathology and Molecular Medicine, 1280 Main Street West, Hamilton, Ontario, Canada L8S4K1.
Methods. 2011 Oct;55(2):114-21. doi: 10.1016/j.ymeth.2011.09.022. Epub 2011 Oct 4.
Evidence from clinical and epidemiological studies indicates that women are disproportionately susceptible to sexually transmitted viral infections. To understand the underlying biological basis for this increased susceptibility, more studies are needed to examine the acute events in the female reproductive tract following exposure to viruses during sexual transmission. The epithelial lining of the female reproductive tract is the primary barrier that sexually transmitted viruses, such as HIV-1 and HSV-2 need to infect or traverse, in order to initiate and establish productive infection. We have established an ex-vivo primary culture system to grow genital epithelial cells from upper reproductive tract tissues of women. Using these cultures, we have extensively examined the interactions between epithelial cells of the female genital tract and HSV-2 and HIV-1. In this review, we describe in detail the experimental protocol to grow these cultures, monitor their differentiation and inoculate with HSV-2 and HIV-1. Prospective use of these cultures to re-create the microenvironment in the reproductive tract is discussed.
来自临床和流行病学研究的证据表明,女性易感染性传播病毒感染的比例不成比例。为了了解这种易感性增加的潜在生物学基础,需要更多的研究来检查女性生殖道在性传播过程中暴露于病毒后发生的急性事件。女性生殖道的上皮衬里是性传播病毒(如 HIV-1 和 HSV-2)感染或穿透所必需的主要屏障,以便开始并建立有性感染。我们已经建立了一种从女性上生殖道组织中生长生殖上皮细胞的体外原代培养系统。使用这些培养物,我们广泛研究了女性生殖道上皮细胞与 HSV-2 和 HIV-1 之间的相互作用。在这篇综述中,我们详细描述了培养这些细胞的实验方案,监测它们的分化,并接种 HSV-2 和 HIV-1。讨论了这些培养物在重建生殖道微环境中的预期用途。