Lee Wai-Ming, Chen Yin, Wang Wensheng, Mosser Anne
Biological Mimetics Inc., 124 Byte Drive, Frederick, MD, 21702, USA.
Methods Mol Biol. 2015;1221:49-61. doi: 10.1007/978-1-4939-1571-2_5.
HeLa cell culture is the most widely used system for in vitro studies of the basic biology of human rhinovirus (HRV). It is also useful for making sufficient quantities of virus for experiments that require highly concentrated and purified virus. This chapter describes the protocols for producing a large amount of HeLa cells in suspension culture, using these cells to grow a large quantity of virus of HeLa-adapted HRV-A and -B serotypes, and making highly concentrated virus stock and highly purified virions. These purified HRV virions are free of cellular components and suitable for experiments that are sensitive to cellular contaminations.
HeLa细胞培养是用于体外研究人鼻病毒(HRV)基础生物学的最广泛使用的系统。它对于为需要高浓度和纯化病毒的实验制备足够量的病毒也很有用。本章描述了在悬浮培养中大量生产HeLa细胞的方案,使用这些细胞培养大量适应HeLa的HRV - A和 - B血清型病毒,并制备高浓度病毒储备液和高度纯化的病毒粒子。这些纯化的HRV病毒粒子不含细胞成分,适用于对细胞污染敏感的实验。