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Isolation and Characterization of Virus-resistant Mouse Embryo Fibroblasts

Department of Biochemistry, University of Leicester, Leicester, England

C. Colby, Jun. and Judith L. N. Hulse

Microbiology Section, University of Connecticut, Storrs, Conn. 06268, U.S.A.

A method is described for the isolation of virus-resistant mutants of mammalian cells in culture. Cells of a mouse embryo fibroblast line (3T6) were treated with a mutagen and subjected to selection for virus resistance. The cells were infected with a temperature-sensitive mutant of Sindbis virus which was allowed to proceed through one cycle of replication at the permissive temperature. On transfer of the cells to the non-permissive temperature, further cycles of virus replication were prevented and non-infected cells allowed to grow out. Repetition of this procedure over 2 months resulted in the isolation of a population of 3T6 cells which resisted infection by Sindbis virus.

Single cell clones were isolated that were resistant to the growth of Sindbis virus and also to the growth of Mengo virus, Semliki Forest virus, vesicular stomatitis virus, Newcastle Disease virus, vaccinia virus and herpes simplex virus.

Received 29 January 1973; accepted 3 May 1973.

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