Department of Pathology, Center for Cell Reprogramming, Georgetown University Medical Center, Washington, DC.
Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University Medical Center, Washington, DC.
J Med Virol. 2020 Nov;92(11):2440-2452. doi: 10.1002/jmv.26093. Epub 2020 Jun 16.
Conventional cancer and transformed cell lines are widely used in cancer biology and other fields within biology. These cells usually have abnormalities from the original tumor itself, but may also develop abnormalities due to genetic manipulation, or genetic and epigenetic changes during long-term passages. Primary cultures may maintain lineage functions as the original tissue types, yet they have a very limited life span or population doubling time because of the nature of cellular senescence. Primary cultures usually have very low yields, and the high variability from any original tissue specimens, largely limiting their applications in research. Animal models are often used for studies of virus infections, disease modeling, development of antiviral drugs, and vaccines. Human viruses often need a series of passages in vivo to adapt to the host environment because of variable receptors on the cell surface and may have intracellular restrictions from the cell types or host species. Here, we describe a long-term cell culture system, conditionally reprogrammed cells (CRCs), and its applications in modeling human viral diseases and drug discovery. Using feeder layer coculture in presence of Y-27632 (conditional reprogramming, CR), CRCs can be obtained and rapidly propagated from surgical specimens, core or needle biopsies, and other minimally invasive or noninvasive specimens, for example, nasal cavity brushing. CRCs preserve their lineage functions and provide biologically relevant and physiological conditions, which are suitable for studies of viral entry and replication, innate immune responses of host cells, and discovery of antiviral drugs. In this review, we summarize the applications of CR technology in modeling host-virus interactions and human viral diseases including severe acute respiratory syndrome coronavirus-2 and coronavirus disease-2019, and antiviral discovery.
传统的癌细胞系和转化细胞系广泛应用于癌症生物学和生物学的其他领域。这些细胞通常与原始肿瘤本身存在异常,但由于遗传操作或长期传代过程中的遗传和表观遗传变化,也可能发生异常。原代培养物可能保留原始组织类型的谱系功能,但由于细胞衰老的性质,它们的寿命或倍增时间非常有限。原代培养物通常产量非常低,而且由于任何原始组织标本的高度变异性,在很大程度上限制了它们在研究中的应用。动物模型常用于病毒感染、疾病建模、抗病毒药物和疫苗的开发研究。由于细胞表面的可变受体,人类病毒通常需要在体内进行一系列传代以适应宿主环境,并且可能受到细胞类型或宿主物种的细胞内限制。在这里,我们描述了一种长期的细胞培养系统,即条件重编程细胞(CRC),及其在模拟人类病毒疾病和药物发现中的应用。使用饲养层共培养(存在 Y-27632 的条件重编程,CR),可以从手术标本、核心或针吸活检以及其他微创或非侵入性标本(例如鼻腔刷洗)中获得 CRC 并快速繁殖。CRC 保留其谱系功能,并提供生物学相关和生理条件,非常适合研究病毒进入和复制、宿主细胞的先天免疫反应以及抗病毒药物的发现。在这篇综述中,我们总结了 CR 技术在模拟宿主-病毒相互作用和人类病毒疾病(包括严重急性呼吸综合征冠状病毒 2 和 2019 年冠状病毒病)以及抗病毒药物发现中的应用。