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This unit describes the maintenance of cell lines used with vaccinia virus, both in monolayer cultures and in suspension. The suspended cell culture is then used in the preparation of vaccinia virus stocks. The preparation of chick embryo fibroblasts (CEF) is also presented for use in the production of the highly attenuated and host range-restricted modified vaccinia virus Ankara (MVA) strain of vaccinia virus. Additionally, support protocols are presented for the titration of standard and MVA vaccinia virus stocks.

Original publication




Journal article


Current protocols in molecular biology

Publication Date



Chapter 16


National Institutes of Allergy and Infectious Diseases, Bethesda, Maryland, USA.


Cells, Cultured, Cell Line, Hela Cells, Fibroblasts, Chick Embryo, Animals, Humans, Vaccinia virus, Cell Culture Techniques, Viral Plaque Assay