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Isolation of temperature sensitive mammalian cells by selective detachment
Author(s) -
Roscoe D. H.,
Read Moira,
Robinson Hildred
Publication year - 1973
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1040820302
Subject(s) - reversion , chinese hamster , hamster , mutant , isolation (microbiology) , biology , population , transformation (genetics) , agar , substrate (aquarium) , microbiology and biotechnology , chemistry , cell culture , biochemistry , bacteria , genetics , gene , phenotype , medicine , environmental health , ecology
Temperature sensitive cells have been isclated from Syrian and Chinese hamster cells using a method based on selective detachment from a glass substrate. The Syrian hamster isolates occurred at a high frequency (about 1 in 10 3 ) and reverted rapidly; polyoma virus transformation conferred on cells the ability to grow, perhaps abnormally, in agar suspension. A slightly modified isolation technique was applied to Chinese hamster cultures and resulted in the isolation of at least one mutant (from a starting population of 5 × 10 8 cells) with a spontaneous reversion rate of less than one in 6 × 10 7 . Treatment of the mutant with ethyl methane sulphonate induced reversion. It was concluded that selective detachment provided a useful method for the isolation of conditional lethal mutants of mammalian cells.