US4302535A
Expired / Lapsed
US
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Assay for mutagenesis in heterozygous diploid human lymphoblasts
Assignee: Massachusetts Institute of Technology
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Filing: 1979-09-27
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Granted: 1981-11-24
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Expires: 1999-09-27
Abstract
Abstract An assay is disclosed for determining mutagenic damage caused by the administration of a known or suspected mutagen to diploid human lymphoblastoid cell lines. The gene locus employed for this assay is the gene for thymidine kinase, uridine kinase, or cytidine deaminase. Since human lymphoblastoid cells contain two genes for these enzymes, heterozygotes of human lymphoblastoid cells are used in this assay.
Inventors
- 👤 Thomas R. Skopek
- 👤 Howard L. Liber
- 👤 Bruce W. Penman
- 👤 William G. Thilly
- 👤 Henry Hoppe, IV
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 21:50
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/5005; G01N33/5008; G01N33/5014; G01N33/5017
Claims Analysis
Total Claims: 8
Claim 1
Independent Claim
142 words
An assay for determining mutagenesis, comprising: a. exposing a culture of human diploid or near-diploid cells capable of continuous division in suspension culture, said cells being heterozygotes having only one active gene for an enzyme for converting a pyrimidine analog to a toxic material for said cells to an agent to be tested for its mutagenic effects on said cells; b. incubating exposed cells for a number of generations to allow full expression of phenotypically-developed resistance to a pyrimidine analog which acts as a substrate for said enzyme and is normally toxic to said cells; c. culturing cells having the full phenotypically developed resistance both in the presence of and without a pyrimidine analog which is toxic to non-mutant cells; and, d. comparing the amount of mutant cells to the amount of non-mutant cells to thereby determine the degree of induced mutagenesis.
Claim 2
Dependent Claim
142 words
An assay for determining mutagenesis, comprising: a. exposing a culture of human diploid or near-diploid cells capable of continuous division in suspension culture, said cells being heterozygotes having only one active gene for an enzyme for converting a pyrimidine analog to a toxic material for said cells to an agent to be tested for its mutagenic effects on said cells; b. incubating exposed cells for a number of generations to allow full expression of phenotypically-developed resistance to a pyrimidine analog which acts as a substrate for said enzyme and is normally toxic to said cells; c. culturing cells having the full phenotypically developed resistance both in the presence of and without a pyrimidine analog which is toxic to non-mutant cells; and, d. comparing the amount of mutant cells to the amount of non-mutant cells to thereby determine the degree of induced mutagenesis.
Claim 3
Dependent Claim
18 words
An assay of claim 1 wherein the active gene codes for thymidine kinase, uridine kinase or cytidine deaminase.
Claim 4
Dependent Claim
15 words
An assay of claim 2 wherein said cells comprise a heterozygote of human diploid lymphoblasts.
Claim 5
Dependent Claim
14 words
A method of claim 3 wherein said pyrimidine analog comprises a 5-halodeoxyuridine or 5-halothymidine.
Claim 6
Dependent Claim
17 words
An assay of claim 4 wherein an active drug-metabolizing system is also added to the cell culture.
Claim 7
Dependent Claim
13 words
An assay of claims 1 or 5 wherein the pyrimidine analog comprises trifluorothymidine.
Claim 8
Dependent Claim
13 words
An assay of claim 6 wherein the active gene codes for thymidine kinase.
References Cited (Backward Citations)
US4066510A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1984-11-23 | 1985-05-25 | 1985-11-24 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1988-11-23 | 1989-05-25 | 1989-11-24 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1992-11-23 | 1993-05-25 | 1993-11-24 | $7700.00 | Window Closed / Lapsed |