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Title: Method for rapid isolation of sensitive mutants

Abstract

Sensitive mammalian cell mutants are rapidly isolated using flow cytometry. A first population of clonal spheroids is established to contain both normal and mutant cells. The population may be naturally occurring or may arise from mutagenized cells. The first population is then flow sorted by size to obtain a second population of clonal spheroids of a first uniform size. The second population is then exposed to a DNA-damaging agent that is being investigated. The exposed second population is placed in a growth medium to form a third population of clonal spheroids comprising spheroids of increased size from the mammalian cells that are resistant to the DNA-damaging agent and spheroids of substantially the first uniform size formed from the mammalian cells that are sensitive to the DNA-damaging agent. The third population is not flow sorted to differentiate the spheroids formed from resistant mammalian cells from spheroids formed from sensitive mammalian cells. The spheroids formed from sensitive mammalian cells are now treated to recover viable sensitive cells from which a sensitive cell line can be cloned. 15 figs.

Inventors:
Issue Date:
Research Org.:
University of California
Sponsoring Org.:
USDOE, Washington, DC (United States)
OSTI Identifier:
516930
Patent Number(s):
5,652,098
Application Number:
PAN: 8-261,030
Assignee:
Dept. of Energy, Washington, DC (United States) PTO; SCA: 550300; PA: EDB-97:120938; SN: 97001831595
DOE Contract Number:  
W-7405-ENG-36
Resource Type:
Patent
Resource Relation:
Other Information: PBD: 29 Jul 1997
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; MUTANTS; CELL FLOW SYSTEMS; CYTOLOGICAL TECHNIQUES; MAMMALS; ANIMAL CELLS; DESIGN; SENSITIVITY

Citation Formats

Freyer, J.P. Method for rapid isolation of sensitive mutants. United States: N. p., 1997. Web.
Freyer, J.P. Method for rapid isolation of sensitive mutants. United States.
Freyer, J.P. Tue . "Method for rapid isolation of sensitive mutants". United States.
@article{osti_516930,
title = {Method for rapid isolation of sensitive mutants},
author = {Freyer, J.P.},
abstractNote = {Sensitive mammalian cell mutants are rapidly isolated using flow cytometry. A first population of clonal spheroids is established to contain both normal and mutant cells. The population may be naturally occurring or may arise from mutagenized cells. The first population is then flow sorted by size to obtain a second population of clonal spheroids of a first uniform size. The second population is then exposed to a DNA-damaging agent that is being investigated. The exposed second population is placed in a growth medium to form a third population of clonal spheroids comprising spheroids of increased size from the mammalian cells that are resistant to the DNA-damaging agent and spheroids of substantially the first uniform size formed from the mammalian cells that are sensitive to the DNA-damaging agent. The third population is not flow sorted to differentiate the spheroids formed from resistant mammalian cells from spheroids formed from sensitive mammalian cells. The spheroids formed from sensitive mammalian cells are now treated to recover viable sensitive cells from which a sensitive cell line can be cloned. 15 figs.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1997},
month = {7}
}