Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1994-06-14
1997-07-29
Ketter, James S.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
4351721, C12Q 168
Patent
active
056520980
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.
REFERENCES:
J. P. Freyer et al. "A New Method For Isolating Radiation-Sensitive Mutant Cells," Abstracts of Papers for the Fortieth Annual Meeting of the Radiation Research Society, Salt Lake City, Utah (Mar. 14-18, 1992) P-21-10, p. 86.
James P. Freyer et al., "Use of Xantham Gum To Suspend Large Particles During Flow Cytometric Analysis And Sorting," Cytometry 10, pp. 803-806 (1989).
James P. Freyer et al., "Viable Sorting Of Intact Multicellular Spheroids By Flow Cytometry," Cytometry 8, pp. 427-436 (1987).
P. A. Jeggo et al., "X-Ray-Sensitive Mutants Of Chinese Hamster Ovary Cell Line Isolation And Cross-Sensitivity To Other DNA-Damaging Agents," Mutation Research 112, pp. 313-327 (1983).
M. Z. Zdzienicka et al., "Mutagen-Sensitive Cells Lines Are Obtained With A High Frequency In V79 Chinese Hamster Cells," Mutation Research 178, pp. 235-244 (1987).
Eklund William
Ketter James S.
Moser William R.
The United States of America as represented by the United States
Wilson Ray G.
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