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Appl Environ Microbiol. 1971 November; 22(5): 914-918
Copyright © 1971 American Society for Microbiology. All Rights Reserved.

Propagation and Purification of High-Titer Human Cytomegalovirus

Department of Pathology, Georgetown University School of Medicine, Washington, D.C. 20007
Laboratory of Biology of Viruses, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20014
Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20014
Electro-Nucleonics Laboratories, Inc., Bethesda, Maryland

ABSTRACT

High-titered yields of human cytomegalovirus (CMV), strain AD 169, were produced in WI-38 cells in large roller bottles. Maximum plaque titers were observed by the 4th day after infection at which time infectivity in the medium was 200 times greater than that associated with the cells. Virus released into the medium was recovered by sedimentation in a sucrose gradient in a continuous-flow centrifuge rotor. Maximal viral infectivity was found at a sucrose concentration of 42%, equivalent to a density of 1.18 g/cm³. Deoxyribonucleic acid extracted from these preparations was about 80% viral and 20% cellular as judged by equilibrium centrifugation in cesium chloride density gradients.